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01 October 2010, Volume 14 Issue 40 Previous Issue    Next Issue

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A comparison of various methods for inducing adult bone marrow mesenchymal stem cells differentiation into neuron-like cells Zhang Yu-hong, Han Xiao-gai, Xin Xiu-yin 2010, 14 (40):  7411-7414.  doi: 10.3969/j.issn.1673-8225.2010.40.001 Abstract ( 258 )   PDF (312KB) ( 312 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) could be induced to differentiate into neuron-like cells through a variety of inducers in vitro. Different inducers of cell differentiation rate, cell viability, as well as the rate of nestin-positive cells staining are different. OBJECTIVE: To explore a better inducer by comparing results of BMSCs differentiation into neuron-like cells using multiple inducers. METHODS: The healthy adult bone marrow was taken in the sterile laminar flow room. Lymphocyte separating medium was added for centrifugation, and the albuginea was obtained. Cells were placed in medium containing basic fibroblast growth factor (bFGF), epidermal growth factor, DMEM and 10% fetal bovine serum in 5% CO2 incubator at 37 ℃. The third passage of cells were divided into β-mercaptoethanol + dimethyl sulfoxide group, brain-derived neurotrophic factor (BDNF) + retinoic acid group, glial cell line-derived neurotrophic factor (GDNF) + retinoic acid group and bFGF group. Inducers were added separately. Cell morphology was observed. Cell viability was measured. Nerve cell markers were detected using immunohistochemistry. RESULTS AND CONCLUSION: Cell viability was compared in each group using trypan blue staining. Cell viability was greatest in the GDNF + retinoic acid group (P < 0.05), but lowest in the β-mercaptoethanol + dimethyl sulfoxide group (P < 0.05). No significant difference in cell viability was determined between the BDNF + retinoic acid group and bFGF group (P > 0.05). Positive rate of nerve cell markers was greater in the GDNF + retinoic acid group compared with β-mercaptoethanol + dimethyl sulfoxide group, BDNF + retinoic acid group and bFGF group (P < 0.05). These suggest that adult BMSCS could differentiate into neuron-like cells. GDNF + retinoic acid was the optimal induction which showed higher living cell energy and positive immunoreactivity to nerve markers (P < 0.05). Related Articles | Metrics

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Differentiation of superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells into cardiomyocytes-like cells in vitro induced by 5-azacitidine Liu Yong-hao, Guo Liang, He Mi-chun, Shen Zhen-ya 2010, 14 (40):  7415-7419.  doi: 10.3969/j.issn.1673-8225.2010.40.002 Abstract ( 354 )   PDF (404KB) ( 339 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) induced by various inducers can differentiate into cardiomyocyte-like cells in vitro. The common inducer was 5-azacitidine. Superparamagnetic iron oxide (SPIO) particle has small diameter and can be phagocytized by stem cells. Its paramagnetism can be detected by magnetic resonance imaging (MRI). Thus, SPIO has been an ideal tracer of transplanted stem cells in living bodies. OBJECTIVE: To observe the effects of differentiation of SPIO-labeled stem cells into cardiomyocytes-like cells in vitro induced by 5-azacitidine, and compare with unlabeled stem cells. METHODS: Pig BMSCs were isolated and cultured. In the experimental group, the third passage of BMSCs were marked with SPIO, and then 5-aza was added in vitro for 24 hours. In the control group, BMSCs were induced by 5-azacitidine directly. 2 weeks later, the induced cells were evaluated with desmin, myosin heavy chain, cardiac troponin I and connexin-43 by immunocytochemical staining. Iron particles were detected by Prussian blue staining. RESULTS AND CONCLUSION: Desmin, myosin heavy chain, cardiac troponin I, connexin-43 were expressed in BMSCs induced by 5-azacitidine. Prussian blue staining showed numerous blue-stained iron particles in the cytoplasm of SPIO marked BMSCs. These indicate that SPIO-marked BMSCs can differentiate into cardiomyocytes-like cells under 5-azacitidine induction in vitro. Related Articles | Metrics

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Human bone marrow mesenchymal stem cells labeled with Feridex in vitro Zhong Yue-si, Lin Ji-zong, Ke Ying-ping, Tang Zhao-feng, Xu Jian-liang, Lin Nan, Xu Rui-yun, Deng Mei-hai　 2010, 14 (40):  7420-7424.  doi: 10.3969/j.issn.1673-8225.2010.40.003 Abstract ( 331 )   PDF (741KB) ( 260 )   Save BACKGROUND: Conventional detection methods of stem cell tracking are lack of succession and noninvasion, because most of them need animal organizations to be inspected. OBJECTIVE: To identify the effect of Feridex on human bone marrow mesenchymal stem cells (BMSCs) activities and determine suitable concentration of Feridex and screen sequence of magnetic resonance imaging (MRI) scan, through labeling BMSCs with Feridex ex vivo. METHODS: Passage 3 BMSCs in good condition were labeled with Feridex of different concentrations (100, 50, 25, 12.5 mg/L). The culture was performed for 24-48 hours. Blank control group included Passage 3 BMSCs that were not labeled. Proliferation of BMSCs of the two groups was detected with methyl thiazolyl tetrazolium (MTT), and growth curve was made. The labeling was identified with Prussian blue staining. MRI scan of BMSCs labeled with Feridex was performed to determine the suitable sequence of MRI scan. RESULTS AND CONCLUSION: There was no effect of Feridex with concentration ≤25 mg/L on BMSCs, and the result of labeling with concentration of 25 mg/L was optimal. Feridex with concentration≥ 50 mg/L would obviously inhibit the proliferation of BMSCs. Results of MRI scan revealed that MRI signal of BMSCs Feridex labeled for 24 and 48 hours or the filial Passage degraded in all sequence T1WI, T2WI and T2*WI3, the change of T2*WI was mostly obvious. These results indicate that Feridex can be used for ex vivo labeling of BMSCs for succession research. The effect of Feridex on BMSCs was in concentration dependablity, and concentration of 25 mg/L was optimal. The result of sequence T2*WI of MRI scan was more suitable to trace Feridex-labeled human BMSCs Related Articles | Metrics

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Culture and osteogenic differentiation of different-aged children bone marrow mesenchymal stem cells in autologous serum in vitro Sun Ke, Wang Guo-bing, Zu Ying, Tang Sheng-ping, Yu Wei, Chen Xiao-wen　 2010, 14 (40):  7425-7429.  doi: 10.3969/j.issn.1673-8225.2010.40.004 Abstract ( 274 )   PDF (462KB) ( 308 )   Save BACKGROUND: Most present studies emphasized on the induced osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) isolated from adults in vitro, but little was known about the osteogenic differentiation of hBMSCs isolated from children with different ages, or its biological characteristics after induced with autologous serum and osteogenic differentiation in vitro. OBJECTIVE: To study culture conditions and methods of different-aged children BMSCs with autologous serum of in vitro culture, and explore its basic biological characteristics after osteogenic differentiation. METHODS: Bone suspension of children, which were divided into three groups according to their age, was collected under aseptic conditions. hBMSCs were isolated and cultivated with media containing autologous serum. The cell growth curve was determined, and expression of cell surface antigens was analyzed by flow cytometry. After differentiation induced by β-sodium glycerophosphate, antiscorbic acid and dexamethasone, formation of alkaline phosphatase, type I collagen and calcium nodules of sub-cultivation cells were accessed by immunohistochemical method. The differentiation efficiency was evaluated. Growth conditions and morphological characteristics of primary cells, passage cells and differentiated cells were monitored using an inverted phase contrast microscope day by day. RESULTS AND CONCLUSION: Among surface antigens cultivated hBMSCs from the three groups, CD29 and CD44 were detected to be positive, while CD34, CD45, CD105, CD106 and HLA-DR were found to be all negative. No significant difference of expression intensities were observed among three groups. The growth rate and differentiation efficiency of children BMSCs decreased with aging. Comparative studies on growth characteristics, surface antigen expression, differentiation potential of children hBMSCs suggested that autologous serum is a safe and effective culture method applicable to clinic with a high differentiation efficiency at the low child ages. Related Articles | Metrics

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Survival and growth of human bone marrow mesenchymal stem cells in different infusion solutions Li Fang, Dong Li-yuan, Chen Yan, Zhang Jian-lin, Niu Bo 2010, 14 (40):  7430-7434.  doi: 10.3969/j.issn.1673-8225.2010.40.005 Abstract ( 303 )   PDF (637KB) ( 334 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) transplantation technology has developed rapidly and the number of patients requiring remote transplantation is increasing gradually. However, the study on cell preservation and transportation is currently inadequate. OBJECTIVE: To compare the effects of common infusion solutions and the infusion solution prepared by our lab on the survival and growth of MSCs. METHODS: Bone marrow of healthy volunteer was extracted to isolate and culture human bone marrow mesenchymal stem cells (hBMSCs) by density gradient centrifugation and adherence screening methods. Cell morphology, cell surface markers and multi-directional differentiation potential were observed and identified. The third-generation hBMSCs were preserved using 10% fetal bovine serum-Dulbecco’s modified Eagle’s medium (FBS-DMEM), the infusion solution prepared by our lab and 5 kinds of common infusion solutions (9 g/L sodium chloride injection, 50 g/L glucose injection, 50 g/L glucose and sodium chloride injection, 9 g/L sodium chloride injection containing 2% human albumin, 9 g/L sodium chloride injection containing 5% human albumin)at  25 ℃ for 1, 2, 4 and 6 hours, cell survival rate were determined. Then, the third-generation hBMSCs were preserved using the infusion solution prepared by our lab at 4 ℃ for 2, 4, 8, 12 and 24 hours. Cell survival rate and cell growth curve were determined. RESULTS AND CONCLUSION: The third-generation hBMSCs were uniform long spindle, closely arranged parallel or swirling. hBMSCs highly expressed CD44 and CD105, and were negative for CD45. After 3 weeks of osteogenic induction, long-spindle BMSCs became polygonal, and von Kossa staining showed the typical black calcified nodules. After 2 weeks of adipogenic induction, cells became short spindle or oval, and oil red O staining showed dense red-stained lipid droplets. At 25 ℃, the survival rate of hBMSCs stored in the infusion solution prepared by our lab was much higher than hBMSCs stored in 5 kinds of common infusion solutions mentioned above. At 4 ℃, the survival and growth ability of hBMSCs stored in the infusion solution prepared by our lab and 10% FBS-DMEM were identical essentially. Results suggested that the infusion solution prepared by our lab according to cell culture medium is a more appropriate solution for maintaining cell activity and extending the extension of cell survival. Related Articles | Metrics

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Feasibility of in vitro CM-Dil-labeled human umbilical cord mesenchymal stem cells passage tracing Chen Li, Wu Ben-qing, Cheng Han-rong, Huang Jin-jie, Ding Lu, Song Jin-zhi 2010, 14 (40):  7435-7438.  doi: 10.3969/j.issn.1673-8225.2010.40.006 Abstract ( 306 )   PDF (377KB) ( 453 )   Save BACKGROUND: The key to study biological characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) is to master their tracing method of hUCMSCs transplantation. OBJECTIVE: To analyze the tracing feasibility of CM-Dil labeled hUCMSCs in vitro. METHODS: hUCMSCs were isolated and cultured in vitro after enzyme digestion. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified in vitro. The fifth passage of cells were labeled with CM-Dil, passaged and observed by fluorescence microscope in vitro.   RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for CD34 and CD40. Cells in G0/G1 phase accounted for more than 80%. Following adipogenic and osteogenic induction, oil red and alkaline phosphatase staining were positive respectively. The efficiency of CM-DiI labeling hUCMSCs was above 90%. Fluorescence intensity gradually decreased in vitro. After passaged for 8 generations, it nearly quenched. These indicated that hUCMSCs have great potential to proliferation and differentiation. The tracing method of CM-DiI labeling hUCMSCs is effective and easy. Related Articles | Metrics

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Potential of human umbilical cord mesenchymal stem cells differentiating into osteoblasts Zhala Ga-hu, Chen Li, Lan Xiao-xia, Chen Xiao, Chen Xiao-yi 2010, 14 (40):  7439-7442.  doi: 10.3969/j.issn.1673-8225.2010.40.007 Abstract ( 297 )   PDF (385KB) ( 355 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (UCMSCs) were rich and more primordial, had strong differentiation ability and low immunogenicity, suitable for transplanting to different individuals. Therefore, UCMSCs were an ideal target cells for cell therapy. OBJECTIVE: To study the isolation and culture of UCMSCs and to explore the potential of osteogenesis differentiation in vitro.  METHODS: UCMSCs were sterilely isolated and cultured. The third passage of cells were divided into induction group and control group. The induction group was treated with ossify induction medium, and control group was treated with stem cell culture medium. The cell morphology was observed under an inverted light microscope. Cell proliferation was determined by MTT assay. The cell viability was detected by double immunofluorescent staining. Cell cycle and cell surface marker were determined using flow cytometry. Following induction, alkaline phosphatase (ALP) activity was measured utilizing microplate reader. Calcium deposition was analyzed using Von Kossa staining. mRNA expression of osteopontin (OPN), ALP and bone sialoprotein (BSP) was measured by reverse transplantation-polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: The passage cells were steady in the cell shapes, with good viability, and highly expressed CD44. Following induction, the cells were positive in von Kossa staining. ALP activity was higher in induction group compared with control group (P < 0.05). The ALP activity was greatest on day 21 (P < 0.05). RT-PCR indicated that ALP mRNA expression was higher in induction group compared with control group (P < 0.05). The BSP and OPN genes were expressed in induction group, which suggested that human UCMSCs can differentiate into osteoblasts. Related Articles | Metrics

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Cell transplantation for the treatment of advanced-stage spinal cord injury: A Meta analysis Deng Zhou-ming, Cai Lin, Zhang Yi, Liu Zheng-jie, Chen Zhi-long, Yang Biao 2010, 14 (40):  7443-7446.  doi: 10.3969/j.issn.1673-8225.2010.40.008 Abstract ( 383 )   PDF (241KB) ( 332 )   Save BACKGROUND: Neural regeneration and functional recovery following spinal cord injury (SCI) is still a hot focus and difficulty in the medical circle. Recently, cell transplantation as a new therapeutic method with deep research has obtained study outcomes, but its clinical curative effect remains controversial. OBJECTIVE: To evaluate the effectiveness of cell transplantation for the treatment of advanced-stage SCI by meta-analyses.  METHODS: Cochrane Database (Issue 4, 2010), PubMed (1966-2010), EMbase (1989-2010), Chinese Biomedical Literature Database (1978-2010), Wanfang Database (1982-2010) were searched to identify the literatures. All studies about cell transplantation for the treatment of advanced-stage SCI were adopted; the quality of researches was critically appraised and data were extracted by 2 investigators independently. It was collected that the patients’ American Spinal Injury Association scores of pre-operation and post-operation, including motor score, light touch and pinprick. RESULTS AND CONCLUSION: Seven cohort studies involving 388 patients were collected. The results of Meta-analyses showed that the difference of American Spinal Injury Association scores between pre-operation and post-operation had statistical significance (P < 0.000 01). The mean increase in ASIA motor scores was 6.14 [95%CI (3.43, 8.85), P < 0.000 01], and the mean increase for light touch was 8.63 [95%CI (5.61, 11.66), P < 0.000 01] and the mean increase for pinprick was 10.65 [95%CI (7.58, 13.72), P < 0.000 01]. There were no severely adverse effects after cell transplantation. Cell transplantation could be a treatment method for advanced-stage SCI, but the effectiveness and safety required being proved by the randomized controlled trials of large sample. Related Articles | Metrics

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Dose-depended effects of bovine pituitary extract, basic fibroblast growth factor and Forskolin on olfactory ensheathing cell proliferation Yang Bo-yu, Wang Feng, Wang Wei 2010, 14 (40):  7447-7452.  doi: 10.3969/j.issn.1673-8225.2010.40.009 Abstract ( 256 )   PDF (375KB) ( 386 )   Save BACKGROUND: Growth factors participate in regulating proliferation, differentiation and metabolism of olfactory ensheathing cells (OECs). Therefore, reasonable sequential stimulation form can be used to reach multiple amplification of cell number. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF), bovine pituitary extract and Forskolin on OECs proliferation as well as dose-effect relationship. METHODS: The OECs were isolated from the neonatal rat olfactory bulb using primary culture technique. The OECs were purified using differential velocity adherent + chemicals + trypsin digestion method. The bFGF, bovine pituitary extract and Forskolin alone and their combination were added. MTT assay was utilized to detect OEC growth in each group. The purity of the OECs was evaluated and growth rate of OECs was calculated using NGFR p75 immunocytochemistry. RESULTS AND CONCLUSION: Following primary culture and purification of OECs, bovine pituitary extract could facilitate OECs growth, but typical dose-effect relationship was not found. bFGF could facilitate OECs growth, which presented typical dose-effect relationship. An additional Forskolin could not promote OEC proliferation. These indicated that combined use of bFGF and Forskolin in a proper concentration significantly enhanced cell proliferation. Results suggest that sequent use of combined growth factors in a proper concentration would be a reasonable strategy to effectively enhance cell proliferation and this might be an ideal way in dealing with seed cell scarcity for treatment with spinal cord injury. Related Articles | Metrics

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Effects of human bone marrow mesenchymal stem cell transplantation on cognitive ability and hippocampus ultrastructure in Alzheimer’s disease rats He Wei, Bo Hai, Mu Xin-hong, Zhang Ling, Li Hai-sheng　 2010, 14 (40):  7453-7457.  doi: 10.3969/j.issn.1673-8225.2010.40.010 Abstract ( 239 )   PDF (556KB) ( 281 )   Save BACKGROUND: There is no effective method to treat Alzheimer’s disease, due to unclear onset mechanism. In clinic, drug therapy is commonly used, but replacement therapy of bone marrow mesenchymal stem cells (BMSCs) remains in basic research stage. There is no report concerning effects of BMSC transplantation in the hippocampus on cognitive ability of Alzheimer’s disease. OBJECTIVE: To investigate the effects of BMSC transplantation on cognitive ability and hippocampus ultrastructure in Alzheimer’s disease rats. METHODS: A total of 30 senile male Wistar rats with Alzheimer’s disease were chosen to prepare models of natural senescence Alzheimer’s disease. Following model establishment, the rats were assigned to three groups. Bilateral hippocampi were selected as transplantation region. In the differentiation and transplantation group, rats received injection of neuronal induced BMSCs 4 μL (2×105 cells). In the BMSC transplantation group, rats received an equal volume of human BMSCs. In the model group, rats were injected bilaterally with physiological saline into the hippocampus. The learning and memory ability of the rats were detected by Y type maze test. The hippocampus ultrastructure was observed with transmission electron microscopy. RESULTS AND CONCLUSION: The learning and memory scores significantly decreased in model group (P < 0.01), increased in BMSC transplantation group (P > 0.05), and significantly increased in the differentiation and transplantation group (P < 0.01). On week 12, compared with model group, the learning and memory scores were significantly higher in BMSC transplantation group and differentiation and transplantation group (P < 0.01). With the electron microscopy, hippocampus neurons were obviously injured in model group, but the majority of neurons were injured mildly in BMSC transplantation group, which the majority of them were almost normal in differentiation and transplantation group. These indicated that BMSC transplantation may improve the cognitive ability of Alzheimer’s disease rats, and neuronal induced BMSC transplantation is superior to undifferentiated BMSCs. Lowering hippocampus neuronal necrosis may be one of mechanisms that BMSCs improve cognitive dysfunction in Alzheimer’s disease rats. Related Articles | Metrics

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Intravenous transplantation of human adipose-derived mesenchymal stem cells for the treatment of spinal cord injury: Correlation factor and behavior evaluation Qiao Xiao-jun, Yang Bo, Guan Fang-xia, Hu Xiang, Du Ying, Zhang Tian-xiang, Tian Yi, Ba Yun-tao, Duan Xiao-bing, Deng Xiao-hui, Gu Chen-xi, Lei Ning-jing, Wang Xiao-wei 2010, 14 (40):  7458-7461.  doi: 10.3969/j.issn.1673-8225.2010.40.011 Abstract ( 379 )   PDF (754KB) ( 390 )   Save BACKGROUND: As a convenience derived stem cells, adipose-derived mesenchymal stem cells (ADMSCs) have been confirmed that it can be differentiated into neuron-like cells after induced with inducer like Danshen root in vitro. Therefore, ADMSCs have the possibility to become a new seed cell which can treat spinal cord injury. OBJECTIVE: To explore the effects of tail vein transplantation with ADMSCs on rat behaviors of acute closed spinal cord injury and expressions of various factors in spinal cord tissue. METHODS: Human ADMSCs were isolated and cultured in vitro under sterile conditions. When ADMSCs were transmitted into the fourth generation, the cell concentration was adjusted to 1×109/L. Model of spinal cord injury was established in rats of saline control and cell transplantation groups. At 1 week following model induction, 1 mL stem cell suspension was injected into the tail vein of rats from the cell transplantation group. The same volume of saline was injected into the saline control group. No treatment was given in the model control group. RESULTS AND CONCLUSION: Compared with the model control group and saline control group, the motor function of the hind limb was significantly recovered in the cell transplantation, but BBB score was significantly increased (P < 0.05). Expression of glial fibrillary acidic protein was significantly decreased (P < 0.05). Positive expression of neuron specific enolase and nestin was significantly increased (P < 0.05). At 3 days and 1 week following transplantation, fluorochrome-labeled ADMSCs were visible in the damaged region and surrounding segments, mainly in 1 cm range of damaged spinal cord segment, showing uneven distribution. These indicated that after the rats which suffered from acute closed spinal cord injury were transplanted via the tail vein with ADMSCs, their behaviors were improved, and the differentiations of neuronal cells were significantly increased in local damaged regions, resulting in rapid repair. Related Articles | Metrics

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Rolipram combined with oligodendrocyte precursor cell transplantation for treatment of spinal cord injury in rats Qi Quan, Bi Zheng-gang, Yang Wei-liang, Li Xue-zhao, Yang Xin, Zhang Lei, Pan Shang-ha 2010, 14 (40):  7462-7469.  doi: 10.3969/j.issn.1673-8225.2010.40.012 Abstract ( 229 )   PDF (637KB) ( 373 )   Save BACKGROUND: There are many reasons for the difficult regeneration of the injured spinal cord axon, and a comprehensive application of several methods will be more effective. OBJECTIVE: To observe axonal regeneration and remyelinization using Rolipram combined with oligodendrocyte precursor cells (OPCs) transplantation in rats after spinal cord injury (SCI). METHODS: Wistar rat models of SCI were established using ALLEN method. Alzet osmotic pumps contained Rolipram 200 μL were embed subcutaneouly in Rolipram treatment and combination groups, release 0.5 μL per hour, 2 mg/(kg•d) for 14 days. After one week, 0.012 5 μmol dibutyryl cyclic-cyclic adenosine monophosphate was infused in the injured spinal cord. OPCs were infused to cell transplantation and combination groups. An equal volume of saline was injected in the SCI group. Basso Beattie and Bresnahan score was assessed weekly postoperation until 4 weeks when all rats were killed and spinal cords were made into frozen sections. Hematoxylin-eosin staining and immunofluorescence method were performed. RESULTS AND CONCLUSION: At 4 weeks postoperation, Basso Beattie and Bresnahan score was greater in the Rolipram treatment and combination groups compared with SCI and cell transplantation groups. Immunofluorescence method showed: the expression of NF200 was strong in Rolipram treatment and combination groups. OPCs were alive and positive for myelin basic protein, which was predominant and myelinated nerve fiber increased in latter. These suggest that Rolipram elevated cyclic adenosine monophosphate levels, promoted functional recovery after SCI, especially combined with OPC transplantation. Related Articles | Metrics

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Influence of different infusion methods of human umbilical cord mesenchymal stem cells on acute tubular necrosis Li Fang, Hu Xiang, Zhao Hong-mei, Jia Dan-bing, Dang Zhi-jie 2010, 14 (40):  7470-7473.  doi: 10.3969/j.issn.1673-8225.2010.40.013 Abstract ( 378 )   PDF (362KB) ( 424 )   Save BACKGROUND: Studies have confirmed that exogenous mesenchymal stem cells can migrate and establish in the nephridial tissue following ischemia/reperfusion injury, differentiate into renal tubular epithelial cells, and recover structure and function of the kidney. At present, it is controversial which is better, arterial injection or intravenous injection, for transplanting stem cells. OBJECTIVE: To observe the effects of different transplantation methods of human umbilical cord mesenchymal stem cells on dogs with acute tubular necrosis (ATN).  METHODS: Healthy dog models of ATN were established and randomly divided into two groups. Human umbilical cord derived mesenchymal stem cells were labeled with DAPI. Then the cells were injected through peripheral veins and right femoral artery into model dogs’ body. 24 hours later, histological changes were observed by light microscope, and the survival of human umbilical cord mesenchymal stem cells was evaluated by fluorescent microscope. RESULTS AND CONCLUSION: Under common optical microscope, pathological changes such as swelling, degeneration and necrosis appeared in renal tubular epithelial cells in both groups. The injected human umbilical cord mesenchymal stem cells could survive in the affected kidney. There was no significant difference on fluorescence intensity between two groups (P > 0.05). These indicated that transplanted human umbilical cord mesenchymal stem cells by two pathways can survive in the kidneys. The outcomes were similar in repairing the kidney. Compared with arterial injection, the transplantation of intravenous injection obtained little trauma. Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cells transplantation and growth associated protein-43 expression in brain repair Wang Na, Cao Wen, Liu Guang-yi 2010, 14 (40):  7474-7478.  doi: 10.3969/j.issn.1673-8225.2010.40.014 Abstract ( 296 )   PDF (366KB) ( 307 )   Save BACKGROUND: The bone marrow stromal cells transplantation can promote rat brain functional recovery, and have reparation function in cortex and hippocampal structure injury. These may be associated with cell self-compensation and nerve growth media, or because nervous irritability injury stimulates target tissue cells to secrete various nerve factor expression. OBJECTIVE: To observe the role of bone marrow mesenchymal stem cells (BMSCs) transplantation on cerebral recovery and nerve regeneration after cerebral ischemia injury in rats from the angle of cell biology. METHODS: In accordance with modified Nagasawa’s method, the middle cerebral artery occlusion/reperfusion (MCAO/R) model was established. BMSCs transplantation was performed, followed by running machine sports training and water maze test. Neurobehavioral scores, learning and memory scores were observed. TUNEL method was used to detect the expression of apoptotic neurons in cortex region and in hippocampus region, and immunohistochemical technique was utilized to determine expression change of growth associated protein-43 protein in two districts. RESULTS AND CONCLUSION: In transplantation group, BMSCs expression was significantly increased in the cortical area and hippocampal CA1 region at 16 hours, peaked at 7 days, and the number of differentiated cells was significantly increased. Following transplantation, modified neurological severity score was lower in the transplantation group at 7, 19 and 21 days compared with model group (P < 0.01). The time of platform latency was obviously shortened in the water maze test in the transplantation group compared with model group (P < 0.05). The number of times of crossing the platform was greater in the transplantation group compared with model group (P < 0.05). The number of apoptotic cells peaked at 24 hours following ischemia/reperfusion injury. At 3 days, infarct volume was maximal. At 19 days following reperfusion, growth associated protein-43 expression peaked. These indicate that cerebral ischemia mediated neurologic impairment in rats. BMSCs transplantation contributed to neural regeneration. Up-regulation of growth associated protein-43 expression suppressed neuronal apoptosis, and further promoted the repair of cerebral infarction. Related Articles | Metrics

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Effects of transplantation of X-box-binding protein-1 gene-modified neural stem cells on focal brain ischemia/reperfusion injury in rats Jin Xian-hua, Zheng Sheng-zhe, Lu Lei, Song Lei 2010, 14 (40):  7479-7482.  doi: 10.3969/j.issn.1673-8225.2010.40.015 Abstract ( 338 )   PDF (324KB) ( 288 )   Save BACKGROUND: Neural stem cells (NSCs) transfected with X-box-binding protein-1 (Xbp-1) gene transplantation to the lesion site not only ensures the stability of the Xbp-1 gene, sustains effects of anti-apoptotic role, but also promotes survival and differentiation of NSCs following transplantation. OBJECTIVE: To study the effects of Xbp1 gene on the distribution, differentiation and apoptosis of NSCs transplanted into rat brain ischemic injury site, the anti-apoptosis effect and recovery of nerve function. METHODS: Adult Sprague Dawley rats were selected to establish models of middle cerebral artery occlusion (MCAO). The rats were randomly assigned to four groups: control group (no treatment), phosphate buffered saline (PBS) transplantation group, NSCs transplantation group, Xbp-1-NSCs transplantation group. At 7, 14 and 28 days following treatment, neurological severity score was scored. Brain tissue was obtained to prepare into sections. Immunofluorescence staining was used to observe survival and distribution of NSCs in the brain. On day 28 following transplantation, terminal deoxyribonucleotidyl transferase-mediated biotin- 16-dUTP nick-end labeling assay was used to determine apoptosis of neural cells in the ischemic region. Western blot assay was utilized to detect Bcl-2 expression levels. RESULTS AND CONCLUSION: At 7, 14 and 28 days following transplantation, neurological severity score was significantly lower in the Xbp-1-NSC transplantation group compared with other three groups (P < 0.05). NSCs could successfully migrate to the ischemic regions, survived and differentiated into mature nerve cells. Survival, proliferation and differentiation ability of Xbp-1 gene-modified NSCs was stronger than the common NSCs (P < 0.05). Compared with other three groups, apoptotic number of NSCs in the ischemic regions was significantly reduced, but Bcl-2 levels increased in the Xbp-1-NSCs transplantation group (P < 0.05). These verified that Xbp-1 gene modification can increase NSCs survival and migration ability, obviously decrease neurological severity score in rat models after transplantation by anti-endoplasmic reticulum stress, as well as promote the recovery of neurological function. Related Articles | Metrics

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Effects of olfactory ensheathing cells transplantation on NG2 expression in spinal cord injury rats Wang Guo-yu, He Xi-jing, Yuan Pu-wei, Li Hao-peng, Chang Rui 2010, 14 (40):  7483-7486.  doi: 10.3969/j.issn.1673-8225.2010.40.016 Abstract ( 265 )   PDF (327KB) ( 349 )   Save BACKGROUND: There is no effective therapeutic strategy for spinal cord injury in clinic. Olfactory ensheathing cells (OECs) transplantation for spinal cord injury has obtained great progress. NG2, an important chondroitin sulfate proteoglycan molecule, suppresses axon growth. OBJECTIVE: To explore effects of OECs transplantation on expression of NG2 in spinal cord injury rats, and to analyze action pathway of OECs transplantation in repair of spinal cord injury. METHODS: A total of 112 rats were equally and randomly divided into four groups: blank group, model group, OECs group and DF12 group. In the blank group, rat spinal cord was completely transected at T10, and partially transected at T9 and T11. No other treatment was performed. Animal models of spinal cord injury were established using spinal cord transsection in other three groups. In the OECs group, OECs transplantation was performed. Each transplanted rat received a total of 20 000 cells. In the DF12 group, DF12 culture medium was injected at the same area. At 1, 3, 7, 14, 28, 42 and 56 days following spinal cord injury, NG2 expression was evaluated according to SP kit instructions. RESULTS AND CONCLUSION: In the blank group, NG2 appeared low expression. In the model group and DF12 group, NG2 expression increased at 24 hours. A highest expression of NG2 was present at 7 days. NG2 expression decreased obviously at 4 weeks. It only appeared local expression at 6 and 8 weeks. In the OECs group, moderately NG2 expression was presented on injury region at 1 day; the expression level was lower in the model group and DF12 group at identical time points, but higher than in the blank group. These indicate that NG2 expression was in a lower lever after OECs transplantation; it was inhibited by OECs transplantation. OECs transplantation would eliminate the chemical barrier of NG2，so that the injured nerve could regenerate. It may be one of the mechanisms of OECs transplantation for treating injured spinal cord. Related Articles | Metrics

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Stem cell transplantation in sequence for treatment of Duchenee muscular dystrophy Yang Xiao-feng, Xu Yi-feng, Lü Nai-wu, Zhang Yi-bin, Wang Hong-mei, Lu Yan, Lü Xin, Zhou Jin-xu, Liu Xin-ping 2010, 14 (40):  7487-7492.  doi: 10.3969/j.issn.1673-8225.2010.40.017 Abstract ( 331 )   PDF (1410KB) ( 518 )   Save BACKGROUND: Duchenne muscular dystrophy (DMD), a well-known genetic lethal in humans, cannot be effectively cured at present. Recently, scholars have performed basic research and animal trials using stem cells for treatment of DMD, and also conduct significant clinical studies. OBJECTIVE: To investigate the feasibility and safety of therapy in patients with DMD who received stem cell transplantation in sequence by observing the change of their motor function, muscle cytothesis and regeneration, expression of dystrophin and altering of depletion gene. METHODS: An 8-years old boy with DMD was treated by stem cell transplantation in sequence in May 2009. Exon 13 was depleted by multiplex ligation-dependent probe amplification. Stem cell transplantation in sequence was as follows: First, intravenous umbilical cord mesenchymal stem cell (UC-MSC) transplantation; Second, intramuscular UC-MSC transplantation; finally, monoploid allogene hemopoietic stem cell transplantation. Sero-enzyme, implantation proof of donor HLA antigen, expression of depletion gene and dystrophin of myocyte membrane, motor function were detected after transplantation. RESULTS AND CONCLUSION: By stem cell transplantation in sequence for DMD, depletion gene could be substituted. Dystrophin of myocyte membrane expresses; sero-enzyme steps down significantly; motor function is improved. It could prevent the progression of disease and patient’s condition is to be ameliorated continuously. Related Articles | Metrics

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N-methyl-D-aspartic acid receptor antagonist dose and proliferation of endogenous neural stem cells in the hippocampus of rats with cerebral ischemia/reperfusion Ren Ming-xin, Zhou Li, Li Na-na, Feng Zhi-bo, Zang Wei-dong, Yuan Guo-yan 2010, 14 (40):  7493-7496.  doi: 10.3969/j.issn.1673-8225.2010.40.018 Abstract ( 243 )   PDF (333KB) ( 326 )   Save BACKGROUND: After cerebral ischemia-reperfusion, excessive excitatory amino acids activate N-methyl-D-aspartic acid (NMDA) receptor which repairs nerve cells by the proliferation and differentiation of endogenous neural stem cells (NSCs), but damages nerve cells by intracellular calcium overload at the same time. Controlling NMDA receptor activation may activate endogenous NSCs and minimize damage simultaneously. OBJECTIVE: To investigate the effect of NMDA receptor antagonist MK-801 on endogenous neural stem cell proliferation of reperfused rat hippocampus. METHODS: A total of 54 healthy male Sprague-Dawley rats were randomized into control group (without any treatment), operation group (establish ischemia model), various doses of MK-801 group. Four-vessel occlusion (Pulsinelli-4VO method) was used to establish global cerebral ischemia/reperfusion model. Different doses of MK-801 were injected into lateral ventricle 30 minutes before cerebral ischemia according to 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 mg/kg. Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect nestin protein and mRNA level expression in each group. RESULTS AND CONCLUDION: When MK-801 was under 0.8 mg/kg, no significant differences in the protein and mRNA expression of nestin were detected among groups (P > 0.05), with the presence of high expression. When MK-801 was equal to 0.8 mg/kg, MK-801 significantly suppressed proliferation of endogenous NSCs. When MK-801 was more than 0.8 mg/kg, inhibitory effect was increased with increased concentration. At 1.2 mg/kg, rats developed adverse reaction such as restlessness and ataxia. The nestin mRNA expression results were consistent with the tendency of protein expression, which suggested that MK-801 dose is associated with the repair of nerve function following cerebral infarction. 0.6 mg/kg is a suitable dose during this test. This concentration can make MK-801 reduce the toxic effect of excitatory amino acids on neurons, can protect neural cells, and cannot greatly affect proliferation of endogenous NSCs. Related Articles | Metrics

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Differentiation of bone marrow mesenchymal stem cells into chondrocyte lineage induced by transforming growth factor beta 3 Zhao Ji-dong, Miao Zong-ning, Qian Han-guang, Peng Wei 2010, 14 (40):  7497-7500.  doi: 10.3969/j.issn.1673-8225.2010.40.019 Abstract ( 263 )   PDF (443KB) ( 437 )   Save BACKGROUND: Transforming growth factor β，a polypeptide growth factor, can induce original mesenchymal stem cells (MSCs) differentiation into cartilage tissue during embryogenesis phase. Previous studies have reported that transforming growth factor β3 and basic fibroblast growth factor can contribute to metabolism and proliferation of chondrocytes. OBJECTIVE: To investigate the feasibility of differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocyte lineage by transforming growth factor β3 in vitro. METHODS: Bone marrow was collected from cancellous bone chips during hip joint surgery or during lower limb bones open operation to isolate and culture BMSCs. Its surface antigen was identified. Conditioned medium containing 10% fetal bovine serum and 10 μg/L transforming growth factor β3 was used. Following induction, cells were stained using toluidine blue staining and type II collagen immunohistochemistry. RESULTS AND CONCLUSION: BMSCs were positive for CD73, CD90, CD105, but negative for CD14, CD34, CD45, CD106 and HLA-DR. The morphology of induced cells was obviously changed. The results of toluidine blue staining and type II collagen staining were positive. These indicated that transforming growth factor β3 has the ability to promote BMSCs differentiation into chondrocytes. BMSCs may prove to be a useful source of tissue engineered seed cells. Related Articles | Metrics

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Proliferation and differentiation from neural stem cells into neurons following coculture with olfactory ensheathing cells Wang Xiu-jiao, Sheng Bao-ying, Huang Zuo-yi, Zhang Xiao-mei, Wei Chun-Jie 2010, 14 (40):  7501-7504.  doi: 10.3969/j.issn.1673-8225.2010.40.020 Abstract ( 253 )   PDF (323KB) ( 406 )   Save BACKGROUND: External factors that effect proliferation and differentiation of neural stem cells (NSCs) contain cytokines and microenviroment. Olfactory ensheathing cells (OECs) secrete a variety of cytokines and change the microenvironment of NSCs. OBJECTIVE: To investigate effects of different concentrations of OECs on the proliferation and differentiation of NSCs. METHODS: NSCs and OECs isolated from Wistar rats were cultured in vitro. 5×107/L NSCs were separately cocultured with 1×107/L, 1×109/L, 1×1011/L OECs. Normal control group was set up, without OECs. The proliferation of NSCs was observed under an inverted fluorescent microscope. At 7 days after co-culture, the expression of neuron specific enolase was detected by immunocytochemistry. Ratio of positive cells to total cells was calculated. RESULTS AND CONCLUSION: At 3 days after co-culture, three different concentrations of OECs promoted the proliferation of NSCs. The effects were significant in the NSCs + 1×109/L OECs group, compared with NSCs + 1×107/L OECs group and NSCs + 1×1011/L OECs group. At 7 days after co-culture, the percentage of neurons immunostaining cells was significantly greater in the NSCs + 1×109/L OECs group, which showed significant differences compared with normal control group (P < 0.01). Related Articles | Metrics

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Double layer cells implantation improves retention rate of vascular prosthesis endothelial cells Pei Fei, He Rui, Li Jun-yan, Zhang Li, Chen Xu 2010, 14 (40):  7505-7508.  doi: 10.3969/j.issn.1673-8225.2010.40.021 Abstract ( 295 )   PDF (346KB) ( 483 )   Save BACKGROUND: Previous studies have shown that vascular endothelial cells were seed in grafts to enhance endothelialization rapidly and early, which can improve graft fluency. But the adhesion of seeded endothelial cells is often poor in single-layer. They are easily washed off under the blood flow. OBJECTIVE: To improve cell retention on the graft with dual cell seeding technique, adding a layer of smooth muscle cells (SMCs) between the graft and vascular endothelial cells. METHODS: Rabbit endothelial cells were cultured by standard enzyme separation technique and cell culture technique. Rabbit smooth muscle cells were isolated and cultured using tissue explant method. The lacz and green fluorescent porotein genes were used to transfer endothelial cells and smooth muscle cells. Polytetrafluoroethylene grafts pretreated with fibronectin were seeded with smooth muscle cell followed by endothelial cells in two-layer cells group. Vascular endothelial cells implantation served as controls. Vascular prosthesis was connected to perfusion models for 2 hours. The density of endothelial cells was calculated prior to and following perfusion. In vivo experiment, cell retention rate was measured following vascular prosthesis was inserted into the rabbit for 7 days. RESULTS AND CONCLUSION: 2 hours following perfusion, cell retention rate was (0.39±0.04) in endothelial cells group, and (0.73±0.07) in the two-lay cells group. The cell retention rate was greater in the two-layer cells group compared with endothelial cells group (P < 0.01). 7 days following vascular prosthesis implantation, the cell retention rate was (0.63±0.10) in the endothelial cells group, and (0.95±0.06) in the two-layer cells group (P < 0.05). Results indicated that using smooth muscle cell as a media layer between endothelial cells and graft surface can improve the retention of seeded endothelial cells in vitro and vivo. Related Articles | Metrics

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Ossification induction and phenotype of a special cell in culture process of rat adipose-derived stem cells Wang Fu-ke, Liu Liu, He Xiao-guang, Zhao De-ping, Dai Xiao-ming, Li Yi-song 2010, 14 (40):  7509-7512.  doi: 10.3969/j.issn.1673-8225.2010.40.022 Abstract ( 228 )   PDF (321KB) ( 315 )   Save BACKGROUND: In previous experiment, a type of special cell that mixed fusiform adipose-derived stem cells (ADSCs) was found. A great quantity of polyhedral cells offer nest shape growth and take on cobblestone shape morphology. This is closely related cells to ADSCs, and it is necessary to clarify its nature. OBJECTIVE: To determine the nature of the special cell by analysis of this specific phenotype and the induction of bone formation of these cells. METHODS: Rat ADSCs were isolate and cultured by collagenaseⅠ, cultured by the medium containing fetal bovine serum. The immunofluorescence method was used to observe expressions and distributions of some special surface marker of the polygonal and fibroblastic cells, such as CD34, CD90, CD133 and Flk-1. Cells at passage 2 were obtained and induced by osteoinductive culture medium. Alkaline phosphatase (ALP) and Von Kossa staining were done on day 14 to test the characteristic of ossification. RESULTS AND CONCLUSION:  ①Shape of cells cultured in fetal bovine serum (FBS) showed that a great quantity of polyhedral cells offered nest growth and took on cobblestone morphology, and the fusiform cells were visible. ②The fusiform cells were positive for CD90 and negative for CD34, CD133 and Flk-1; the polyhedral cells were positive for CD34 and Flk-1, and a part of polyhedral cells were positive for CD133, and negative for CD90. ③ In the process of ALP staining, a great quantity of polyhedral cells died, forming blank region; that of some fusiform cells was positive. ④The polyhedral cells died in the process of Von Kossa staining, presenting blank region; a part of fusiform cells formed red calcium nodus. The results have shown that the special cell is vascular endothelial cells in the culture process of rat ADSCs. Related Articles | Metrics

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Differentiation inclination of human embryonic stem cells reversibly affected by different culture systems Luo Shu-wei, Lin Ge, Sun Zheng, Xie Ping-yuan, Lu Guang-xiu 2010, 14 (40):  7513-7518.  doi: 10.3969/j.issn.1673-8225.2010.40.023 Abstract ( 295 )   PDF (664KB) ( 343 )   Save BACKGROUND: Human embryonic stem cells (hESCs) can be maintained in feeder dependent and chemical defined culture systems, and have the capacity to differentiate into three germ layers derived cells. OBJECTIVE: To compare the effects of feeder cells and chemical defined culture system on characteristics of hESCs. METHODS: hESCs in feeder dependent culture system (passage 27) were transferred to chemical defined culture system for 56 passages and returned back to feeder culture system. hESCs in three culture conditions (feeder dependent culture system for 70 passages, chemical defined culture system for 56 passages or the condition hESCs transferred from chemical defined culture system to feeder dependent culture system for 13 passages and 20 passages) were analyzed by flow cytometry for SSEA4. Expression of multipotency gene and differentiation gene in three embryonic layers was determined following hESCs were induced to differentiate under three culture conditions. RESULTS AND CONCLUSION: hESCs exhibited different differentiation inclinations in embryonic bodies conditions in feeder dependent and chemical defined culture systems, and the differentiation inclinations can be converted reciprocally. In chemical defined culture system, hESCs showed anti-differentiation to neural ectoderm cells, and this anti-differentiation can be rescued when hESCs were transferred back to feeder dependent culture system. The gene expression of Nanog may have an important role in embryonic bodies differentiation. At the same time, SSEA4 subpopulations hESCs showed the similar pattern. There may be some relationships between the SSEA4 subpopulations and differentiation inclination in feeder dependent and chemical defined culture systems. Related Articles | Metrics

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Influence of Thy-1.1 stem cell transplantation on nitric oxide synthase of injured artery Liu Hua-dong, Dong Shao-hong, Jiang Xin, Luo Lin-jie 2010, 14 (40):  7519-7523.  doi: 10.3969/j.issn.1673-8225.2010.40.024 Abstract ( 313 )   PDF (814KB) ( 301 )   Save BACKGROUND: At present, effects of stem cell transplantation on restenosis following angiopoiesis are controversial. It is of great significance to explore effects of stem cell transplantation on restenosis and its action mechanism. OBJECTIVE: To study the effect of local transplantation of rat Thy-1.1 stem cells on endothelial hyperplasia following common carotid artery aortic sac, explore effects of stem cell transplantation on endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS), and evaluate the relationship of restenosis and NOS after stem cell transplantation. METHODS: Male 4-6 weeks Sprague Dawley rats were used for preparing Thy-1.1 stem cells. Female Sprague Dawley rats were randomly divided into three groups. The rats of stem cell group were injected about 5×106Thy-1.1 stem cells into the injured artery after carotid artery injury. The rats from injury group underwent carotid artery injury and were injected with the same amount of saline; the uninjured right carotid arteries of the injury group rats were harvested for control group. The animals were killed immediately and 3, 7, 14, 21, 28 days after injury, and the samples of carotid artery were harvested. The intimal thickness was detected using histomorphological methods, and analyzed using computer image analysis. Reverse transcription-polymerase chain reaction was utilized to determine the expression of eNOS mRNA and iNOS. RESULTS AND CONCLUSION: The intimal thickness was thinner in stem cell group compared with injury group (P < 0.05). The eNOS mRNA expression was significantly lower and iNOS mRNA was higher in injury group than in control group (P < 0.05). eNOS mRNA and iNOS mRNA expression was significantly higher in stem cell group compared with injury group (P < 0.05). These indicate that local transplantation of Thy-1.1 stem cells decreased intimal formation and could repair saccule injury, which may be associated with the over-expression of NOS and accelerated re-endothelialization following common carotid artery saccule injury. Related Articles | Metrics

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Establishment of humanized beta-thalassemia mouse models Lai Ying-hui, Lai Yong-rong, Yang Gao-hui, Luo Lin 2010, 14 (40):  7524-7528.  doi: 10.3969/j.issn.1673-8225.2010.40.025 Abstract ( 493 )   PDF (560KB) ( 387 )   Save BACKGROUND: Progress in the gene therapy of β-thalassemia would not have been possible without the development of suitable animal models of the disease. Mouse models are the most useful and are generally adopted as first line in vivo studies. OBJECTIVE: To generate humanized β-thalassemia mice models, and to provide the similar reliable humanized animal models towards the genetic treatment of β-thalassemia. METHODS: Totally 15 nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice were randomly divided into three groups: β- thalassemia/HbE bone marrow transplantation group, β-thalassemia bone marrow transplantation group and blank control group. The bone marrow mononuclear cells of β-thalassemia/HbE and β-thalassemia patients were transplanted into the sublethally irradiated (60Coγ ray 350cGy) NOD/SCID mice of the transplantation group by tail vein injection. RPMI-1640 medium was infused into blank control group. After transplantation, human recombinant growth factors consisting of recombinant human erythropoietin (rhEPO) and recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) were injected into the mice intraperitoneally. The survival status of the mice was recorded. Five weeks after transplantation, bone marrow and peripheral blood were collected to detect human CD45+cell proportion, β-thalassemia gene, to observe α-globin chain precipitates in erythroid cells.The spleen and liver were observed, and iron staining was performed. RESULTS AND CONCLUSION: Five weeks after transplantation, the percentages of human CD45+ cells of the β-thalassemia/HbE bone marrow transplantation group in bone marrow and peripheral blood were 7.21% and 4.08% in +47 day, 7.37% and 6.03% in +61 day, respectively; those of the β-thalassemia bone marrow transplantation group were 8.17% and 3.43% in +36 day, 16.82% and 8.89% in +45 day, respectively. While in blank control group, the percentage of that of each mouse was 0, respectively. The results of β-thalassemia gene mutations corresponded with the donor. There were α-globin chain precipitates in some erythroid cells. The ratios of the spleen length to body weight of the transplantation group were greater than those of blank control group (P < 0.05). The iron deposition was observed in liver and spleen of mice from the transplantation group. Results suggest that there are humanized cell and β-thalassemia features in the mice of β-thalassemia or β-thalassemia/HbE bone marrow transplantation group, and humanized β-thalassemia mice models are generated successfully. Related Articles | Metrics

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Mononuclear cells and platelets isolated from human bone marrow using hydroxyethyl starch by one-step way versus two-step way Ma Yan, Chen Shuang, Bi Xiao-juan, Jiang Ming 2010, 14 (40):  7529-7532.  doi: 10.3969/j.issn.1673-8225.2010.40.026 Abstract ( 340 )   PDF (313KB) ( 311 )   Save BACKGROUND: It can improve the concentrated efficiency of mononuclear cell to use hydroxyethyl starch with lymphocyte separating medium. However, there is no report addressing the residual volume of platelets. OBJECTIVE: To detect the efficiency of isolating mononuclear cells and residual volume of platelets using hydroxyethyl starch by one step or two-step way. METHODS: Bone marrow was obtained from the samples of patients treated with adult autologous bone marrow karyocyte transplantation performed at the First Affiliated Hospital, Xinjiang Medical University, China. Bone marrow tissues acquired from patients were sedimentated by hydroxyethyl starch solution only, or followed by further centrifugation with lymphocyte separating medium. Karyocytes, mononuclear cells and platelet number by these two methods were observed. RESULTS AND CONCLUSION: The average percentage of mononuclear cells in two-step group (89%) was significantly higher than one-step group (45%) (P < 0.05). Moreover, the number of platelets was lower in two-step group (median: 73.00) than one-step group (median: 367.50) (P < 0.05). Results have suggested that the isolation efficiency of using hydroxyethyl starch solution by two-step way from human bone marrow was higher than one-step way. Related Articles | Metrics

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Biological characteristics of placenta-derived mesenchymal stem cells Zhang Hong-yan, Yang Nai-long 2010, 14 (40):  7535-7538.  doi: 10.3969/j.issn.1673-8225.2010.40.028 Abstract ( 283 )   PDF (266KB) ( 573 )   Save BACKGROUND: Recently, the study of mesenchymal stem cells (MSCs) makes some progresses, but there are still two major problems: lack of donor and immune rejection. Placenta-derived mesenchymal stem cells (PMSCs) have many advantages and may become a new source of MSCs. OBJECTIVE: To study the progress in the isolation, culture and induced differentiation of PMSCs. METHODS: A computer-based online search of Pubmed database was undertaken for the related articles published from 1993 to 2009 with the key words of “mesenchymal stem cells, placenta”. 180 literatures were retrieved and primarily checked. Repeated studies were excluded. Of the 41 selected literatures, 3 were reviews and the others were clinical or basic experiment study. RESULTS AND CONCLUSION: Compared to research relatively large number of bone marrow mesenchymal stem cells, PMSCs due to a low immunogenicity, facilitated access to materials, non-invasive procedures, no ethical restrictions, large differentiation potential, proliferative ability, easy preparation and other characteristics of industrialization, become a key source of regenerative medicine and functional stem cells with the prospects for clinical application. Related Articles | Metrics

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Ability and mechanism of stem cells maintaining pluripotency Tian Li-na, Miao Jing-cheng 2010, 14 (40):  7539-7542.  doi: 10.3969/j.issn.1673-8225.2010.40.029 Abstract ( 256 )   PDF (253KB) ( 309 )   Save BACKGROUND: Recent studies have found that adult cells can be reprogrammed and back to embryonic state, which provides possibility for the use of adult stem cells to treat disease. Although there are many researches addressing stem cell proliferation and self-renewal, its mechanism is still little known. OBJECTIVE: To summarize the ability and mechanism of stem cells maintaining pluripotency at home and abroad. METHODS: Pubmed database was searched for the related articles about the mechanism of stem cells maintaining pluripotency published from January 1993 to March 2010. The key words were “stem cells, pluripotency, mechanism”, which were used for selecting some related articles by inputting in title and abstract. The literatures in the same field that published recently or in authorized journals were included. There were 122 articles after the initial survey. Finally, 33 articles were included according to inclusion criteria. RESULTS AND CONCLUSION: Stem cells maintain pluripotency and self-renewal ability by regulating transcription factor expression and epigenetics, as well as microRNAs (miRNAs) effects. The factors such as Oct4, Nanog and Sox2 in regulatory network and epigenetics modification such as histone and DNA methylation and miRNAs exert interactions to inhibit gene expression that promotes stem cell differentiation, which finally forms a regulatory network to maintain stem cell pluripotency and self-renewal. Related Articles | Metrics

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Application prospect of induced pluripotent stem cells Liu Kang, Yi Qiang-ying, Song Gui-qin, Feng Gang 2010, 14 (40):  7543-7546.  doi: 10.3969/j.issn.1673-8225.2010.40.030 Abstract ( 374 )   PDF (274KB) ( 609 )   Save BACKGROUND: Differentiated somatic cells can be reprogrammed into induced pluripotent stem cells (iPS) by transduction of defined transcription factors. Undoubtedly, iPS technology is a great breakthrough of stem cell research in recent years. iPS breaks away from restrictions on source material and ethics. iPS technology elicits a great promise for patient-specific cell therapy and regenerative medicine. OBJECTIVE: To review iPS preparation procedures, limited conditions and mechanisms, collection and application prospect. METHODS: The first author retrieved PubMed database (www.ncbi.nlm.nih.gov/PubMed) for literatures concerning iPS preparation and production mechanisms published from January 2006 to March 2010. The key words were “induced pluripotent stem cells” in English. Simultaneously, some articles were retrieved by hand. Duplicated articles were excluded. Finally, 34 literatures were included. RESULTS AND CONCLUSION: The establishment of iPS mainly contains following procedures: ① choice of recombinant factors; ② choice of target cells; ③ introduction of recombinant factors; ④ expression of recombinant factors in target cells; ⑤ production of iPS; ⑥ identification of recombinant cells. DNA methylation, histone modification and methylation and p53 gene expression play important roles in the process of somatic cells reprogrammed into iPS. The study of iPS technique was still in an early phase, but it has a broad application prospect. In particular, the acquisition of patient-specific and disease-specific iPS provides great cell seed source for understanding the onset mechanism of diseases and for evaluating drug safety. Related Articles | Metrics

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Wnt signaling and differentiation of neural stem cells Bian He-tao, Huang Wei 2010, 14 (40):  7547-7550.  doi: 10.3969/j.issn.1673-8225.2010.40.031 Abstract ( 332 )   PDF (335KB) ( 401 )   Save BACKGROUND: Studies have suggested that neural stem cells (NSCs) transplanted in the host can differentiate into neurons or glial cells. Wnt signaling pathway is closely associated with NSC differentiation. The oriented differentiation of NSCs can be controlled by regulating Wnt signaling pathway. OBJECTIVE: To summarize the relationship of NSC differentiation and Wnt signaling pathway. METHODS: A computer-based online search of Medline, Ovid, CNKI, and EBSCO databases was undertaken for the literatures of NSCs published from February 2002 to March 2010. The key words were “neural stem cells, neural regeneration, Wnt signaling, neuron, differentiation”. The literatures associated with NSC differentiation and with Wnt signaling system were included, whereas repetitive studies were excluded. A total of 30 remaining literatures were used for review. RESULTS AND CONCLUSION: NSCs possess the potential of self-renewal and multi-directional differentiation, can differentiate into many types of cells in the central nervous system. Wnt signaling pathway plays an important role in NSC differentiation. This study analyzed NSCs, Wnt signaling pathway, the correlation of Wnt signaling pathway and NSC differentiation. However, precise mechanism of controlling NSC differentiation by Wnt signaling pathway remains unclear. Related Articles | Metrics

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Characteristics and mechanisms of mesenchymal stem cell culture and induced differentiation　 Zhao Lin, Shen Yan-qing, Rong Chun, Zhou Yan-bing, Xia Chang-suo 2010, 14 (40):  7551-7554.  doi: 10.3969/j.issn.1673-8225.2010.40.032 Abstract ( 360 )   PDF (259KB) ( 373 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) are a class of adult stem cells with multi-directional differentiation potential. OBJECTIVE: To review MSCs separation, identification, characterization and application prospects. METHODS: The Medline database was searched for articles published from January 1996 to March 2010. The key words were “mesenchymal stem cells”. The China National Knowledge Infrastructure database was retrieved for articles published from 1979 to February 2010. The key words were “mesenchymal stem cells, culture, differentiation”. RESULTS AND CONCLUSION: A total of 400 articles concerning MSCs isolation, culture, identification and differentiation were collected, including 86 Chinese articles and 314 English articles. Published earlier, repeated articles and those with similar studies were excluded, and 36 articles in accordance with inclusion criteria were included. We found that there are many methods concerning MSCs isolation, culture and identification, but there is no uniform standard, the mechanism of inducing cell differentiation remains unclear, and there are many problems in application.　 Related Articles | Metrics

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In vitro culture and homing mechanism of bone marrow mesenchymal stem cells Qin Ji-zheng, Han Xiao-dong 2010, 14 (40):  7555-7559.  doi: 10.3969/j.issn.1673-8225.2010.40.033 Abstract ( 293 )   PDF (343KB) ( 408 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have multi-directional differentiation potential, and can subculture in vitro. Under special conditions, multiple factors participate in regulating BMSCs towards damaged tissue organs for repair and reestablishment. OBJECTIVE: To review BMSCs homing mechanism and clinical application in the future. METHODS: A computer-based online search was performed in SCI database for articles published from January 1999 to December 2008. The key words were “bone marrow mesenchymal stem cell, homing, clinical, therapy” in English. Totally 66 articles were retrieved. RESULTS AND CONCLUSION: BMSCs are a kind of stem cells with potential of infinite proliferation and multi-directional differentiation, besides hematopoietic stem cells. Under special induced conditions, BMSCs can homing to a specific tissue and differentiate into corresponding tissue cells, resulting in playing repair and reestablishment effects. In addition, BMSCs are easy to be extracted, with strong amplification ability, no ethical problem or rejection. Therefore, BMSCs have a bright future in the clinical application to many obstinate diseases. Related Articles | Metrics

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Stem cells transplantation and the restoration of sports brain injury Wang Chun-liang, Liu Rui-lian 2010, 14 (40):  7560-7563.  doi: 10.3969/j.issn.1673-8225.2010.40.034 Abstract ( 275 )   PDF (294KB) ( 293 )   Save BACKGROUND: The present study for the head injury, particularly brain injury in sports is one of the hot topics of current research. Stem cells have self-renewal capacity, differentiation potential, and provide broad application prospects for many sports brain injury. OBJECTIVE: By analyzing the movement of the pathological changes of brain injury and stem cell technology, to study application of stem cell technology to brain injury in sports, and to promote their important role in functional recovery. METHODS: Medline database (1994-01/2010-03), Chongqing VIP database (1994-01/2010-06), Tsinghua Tongfang database (1994-01/2010-06) were searched by computer. English key words were “stem cell, spots brain injury, neural stem cells, mesenchymal stem cells”. Chinese key words were “stem cell, spots brain injury, neural stem cells, bone marrow mesenchymal stem cells”. Inclusion criteria: brain injury with exercise and stem cell technology-related literature. Exclusion criteria: repetitive research, old literature. A total of 126 relevant documents were retrieved. Through the study of the repeatability study and the old standard, 23 literatures were included. RESULTS AND CONCLUSION: The study of application of stem cell transplantation to brain injury has obtained great progresses. However, its application to brain injury in sports remains in the exploration stage and requires further investigations. We found that stem cells, especially neural stem cells and bone marrow mesenchymal stem cells for the treatment of brain injury in sports play an important role, and its prospects for more extensive research. Related Articles | Metrics

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Application of induction and differentiation of neural stem cells in the field of sports medicine Zhang Ke-bin, Liu Rui-lian 2010, 14 (40):  7564-7567.  doi: 10.3969/j.issn.1673-8225.2010.40.035 Abstract ( 307 )   PDF (328KB) ( 278 )   Save BACKGROUND: Neural stem cells (NSCs) with their self-renewal, multi-directional differentiation, proliferation, division, injury and disease on the characteristics of strong reaction with the nervous system for a variety of genetic diseases and acquired diseases have brought new hope. OBJECTIVE: To summarize NSC differentiation and its mechanisms, and to explore its application to sports medicine research. METHODS: A computer-based online search was performed in the PubMed database and Chinese journal full-text database for articles published from 1994 to 2010 concerning NSCs and exertional disease. RESULTS AND CONCLUSION: At present, the role of the NSCs in myocardial ischemia, myocardial infarction, neuronal injury, spinal cord injuries and other related diseases research has achieved some achievements, and the application to sports medicine research has broad prospects. Although the NSCs with the diversity of its potential and have the advantage of self-renewal are extensively used in clinic, there are still many problems to be resolved, such as progenitor cells of NSCs and the NSCs to further differentiate into neurons or glial cells whether it has a specific function; in vitro induced NSCs to be transplanted into neural tissue and directional migration, whether they have compatibility of autologous nerve cells in the body and the host, whether they can survive and proliferate for a long term; in vitro-induced NSCs play a role in nerve repair in the integration of autologous neural cells in the host. The research of induced differentiation of NSCs in other sports injuries has not yet been solved. Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation and pravastatin treatment via artery for early femoral head avascular necrosis Li Biao, Liu Jin-song, Wang Kun 2010, 14 (40):  7568-7571.  doi: 10.3969/j.issn.1673-8225.2010.40.036 Abstract ( 199 )   PDF (291KB) ( 322 )   Save BACKGROUND: Stem cell transplantation has recently been used to treat avascular necrosis of the femoral head. The onset of femoral head necrosis is associated with various reasons-induced ischemia in local tissue. Studies have found that the onset of femoral head necrosis may be correlated with decreased function of osteoblasts and bone marrow stromal cells. OBJECTIVE: To investigate the feasibility of autologous bone marrow mesenchymal stem cell (BMSC) transplantation and pravastatin in treatment of avascular necrosis of femoral head in I – II phases via artery. METHODS: A total of 32 patients (49 hips) with avascular necrosis of femoral head were selected. BMSCs were collected following mobilization of colony cell stimulating factors. 15 mL non-ionic contrast media were injected into the affected common iliac artery after successful puncture at the normal femoral artery by Seldinger technique. Autologous stem cell suspension was slowly injected twice through the catheter into the rotation, the lateral artery and obturator arteries, each injection longer than 5 minutes. After the surgery of interventional therapy, oral pravastatin was used. The patients were followed up after the operation. The clinical symptoms of avascular necrosis of femoral head and knee joint activity were evaluated. RESULTS AND CONCLUSION: All 32 patients (49 hips) were followed up over 6 months postoperatively. Hip pain was lessened. Significant improvement in hip function was seen after treatment. The scope of joint activities was significantly expanded, and walking distance was increased. The abduction function was resumed significantly. Angiography of the femoral head in 16 patients (21 hips) at 6 months after treatment revealed that new blood vessels in the femoral head became more, and blood supply was improved. These suggest that BMSC transplantation and pravastatin treatment via artery on early femoral head necrosis have little damage, are well effective in functional recovery, can prevent or delay disease progress. Related Articles | Metrics

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Autologous transplantation of peripheral blood mononuclear cells for lower limb ischemia in 27 cases Li Huan-yu, Hu He-jie, Deng Fu-sheng, Wang Xiao-tian, Sun Xiao-jie 2010, 14 (40):  7572-7575.  doi: 10.3969/j.issn.1673-8225.2010.40.037 Abstract ( 237 )   PDF (343KB) ( 374 )   Save BACKGROUND: The treatment approach of lower limb ischemia from the traditional surgical bypass to now transluminal angioplasty needs better distal arterial outflow tract. Arterial bypass and endovascular treatment are unsuitable for the patients who are lack of arterial outflow tract and cannot bear surgical trauma. OBJECTIVE: To observe the clinical outcome of autologous transplantation of peripheral blood mononuclear cells for treatment of lower limb ischemia. METHODS: A total of 27 patients with lower limb ischemia, because of occluded outflow tract they cannot do bypass grafting and intracavitary angiopoiesis, were enrolled. The patients received subcutaneous injection of recombinant granulocyte colony-stimulating factor 300 μg/d. Following 3 days of mobilization, blood cell separator 3000Plus was used to collect peripheral blood mononuclear cells, which were injected into ischemic limbs gastrocnemius muscle. 3 months later, the improvement in affected limbs was observed. RESULTS AND CONCLUSION: Follow-up was performed from 3 to 60 months, averagely 22 months. Each index was improved in 19 cases (70%), and not improved in 8 cases (30%). Main outcome measures including lower limb skin temperature, rest pain, intermittence claudication distance, insensible feeling, ulcers, ankle-brachial pressure index and transcutaneous oxygen pressure were significantly improved. These indicated that the autologous transplantation of peripheral blood stem cells represents a safe and effective therapeutic approach for lower limb ischemia with occluded outflow tract. Related Articles | Metrics

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Serum levels of alpha-fetoprotein and alpha-fetoprotein variants in decompensated cirrhosis patients following intrahepatic transplantation of peripheral blood stem cells via portal vein Li Nan, Li Na, Shi Yu-ling, Zhu Chao-hui, Sha Li-na, Wu Kai 2010, 14 (40):  7576-7579.  doi: 10.3969/j.issn.1673-8225.2010.40.038 Abstract ( 237 )   PDF (249KB) ( 349 )   Save BACKGROUND: Transplantation of autologous stem cells for treatment of liver cirrhosis has been widely reported. But up to now, there exist some concerns for clinical physicians, including relationship between stem cells and post-transplantation prognosis/turnover of liver cirrhosis, directed differentiation of stem cells in the impaired liver, and malignant phenotype. OBJECTIVE: To dynamically monitor the serum levels of alpha-fetoprotein (AFP) and AFP variants (AFP-L3) in decompensated cirrhosis patients following intrahepatic transplantation of peripheral blood stem cells via portal vein and evaluate the safety of this treatment method. METHODS: A total of 44 decompensated cirrhosis patients who underwent intrahepatic transplantation of peripheral blood stem cells via portal vein in the 309 Hospital of Chinese PLA in April 2007 were included in this study. Prior to and after surgery, serum levels of AFP and AFP-L3 were detected by chemiluminescence. Through the use of a positive criterion for liver cirrhosis, i.e., the proportion of AFP-L3 in AFP [AFP-L3 (%)] ≥10%, and the relationship between decompensated cirrhosis treatment using stem cells transplantation and the malignant phenotype of liver cancer were analyzed. RESULTS AND CONCLUSION: At 2 months after surgery, serum level of AFP showed a transient increase. There was no significant difference in AFP-L3 (%) between prior to and after surgery (P > 0.05). No significant difference in AFP-L3-positve rate, as well as AFP-L3 (%), existed among patients with different serum level of AFP. These findings indicate that clinical symptoms and liver function of decompensated cirrhosis patients recovered to some extent after transplantation of peripheral blood stem cells via portal vein. Results regarding serum level of AFP-L3, a serological marker of liver cancer, did not demonstrate the appearance of malignant biological phenotype. Related Articles | Metrics

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Effects of angelica polysaccharides on the proliferation of mouse skeletal muscle satellite cells in hematopoietic microenvironments in vitro Wang Tao, Feng Li, Wang Xiao-ling, Song Hai-lin 2010, 14 (40):  7580-7582.  doi: 10.3969/j.issn.1673-8225.2010.40.039 Abstract ( 236 )   PDF (253KB) ( 379 )   Save BACKGROUND: It is hopeful that skeletal muscle satellite cells (SMSCs) can be served as seed cells for hematopoietic reconstitution. Angelica polysaccharides (APS) can not only promote hematopoietic stem cells and hematopoietic progenitor cells proliferation and differentiation, but also change the growth characteristics of SMSCs. OBJECTIVE: To investigate the effects of APS on the proliferation of mouse SMSCs in different culture environments. METHODS: SMSCs were procured by a modified method from new born mouse. The α-actin protein of the SMSCs was examined by immunohistochemistry at 5 days after culture. SMSCs were cultured and synchronized for 24 hours in the 96-well plate. After that, SMSCs were assigned into the blank control group, marrow stroma cell supernatant group, APS DMEM/F12 groups (contained 50, 100, 200, 300, 400 mg/L APS) and the marrow stroma cell conditioned medium (disposed by 50, 100, 200, 300, 400 mg/L APS in DMEM/F12). The proliferation of SMSCs was determined by MTT. RESULTS AND CONCLUSION: The α-actin was positive in the cultured SMSCs. MTT results demonstrated that, SMSCs showed a proliferative property in the marrow stroma cell conditioned medium groups. Additionally, the marrow stroma cell conditioned medium can effectively alter growth characteristics of SMSCs in a dose-dependent manner. Related Articles | Metrics

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In vitro female germline potential of human umbilical cord-derived matrix stem cells Li Cai-xia, Wang Feng-ying, Liang Zhi-qing, Li Yu-yan, Yu Chi-yang, Chang Qing, Long Ling 2010, 14 (40):  7583-7587.  doi: 10.3969/j.issn.1673-8225.2010.40.040 Abstract ( 221 )   PDF (322KB) ( 325 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BM-MSCs) have been shown to possess the potential to differentiate into oocytes. However, immune rejection and a limited number of donors of BM-MSCs constrain the applications of BM-MSCs. Several studies have demonstrated that human umbilical cord matrix stem cells (UC-MSCs) also have an intrinsic ability to differentiate into oocyte-like cells in vitro. OBJECTIVE: To establish the method for UC-MSCs culture and to investigate the in vitro differentiation potential of UC-MSCs towards germ cells. METHODS: Umbilical cord from full-term normal deliveries was obtained in sterile condition. Collagenase I-digested cells were cultured in DMEM. The immunophenotype of cells was determined by flow cytometry. Lipoblasts, osteoblasts and chondroblasts were induced in different condition cultures. The expression of germ cells specific marker in UC-MSCs was determined by reverse transcription-polymerase chain reaction. Follicular fluid was employed to induce UC-MSCs differentiation into germ cells. RESULTS AND CONCLUSION: Spindle-like umbilical cord cells were shown and cells in culture were extended to more than 10 passages. BM-MSCs-like immunophenotypes were shown: CD29, CD44, CD73 (SH3), CD90 and CD105 (SH2) were positive; SSEA-4 was weakly positive; CD31, CD34, CD45 and HLA-DR were negative. After UC-MSCs were induced in different condition cultures, lipid droplet-, bone tubercle-, and cartilage tubercle-like structures emerged and the mRNA expressions of specific gene of fat, bone and cartilage were observed. Germ cells markers, OCT4, Stella, Ifitm3, were expressed in UC-MSCs. After induced by 5%, 10% or 20% follicular fluid, cells aggregated and oocyte-like structures were observed. Human UC-MSCs could be cultured and amplificated in vitro. UC-MSCs showed immunophenotypes similar to BM-MSCs. UC-MSCs had the potential to differentiate into lipoblasts, osteoblasts, and chondroblasts. Oocyte-like structure was induced in vitro from UC-MSCs with germ cells specific marker. These findings suggest that UC-NSCs have the potential to differentiate into germ cells. Related Articles | Metrics

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Effects of prenatal pulsed electromagnetic fields on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspring Li Xia, Chen Rui, Jia Ning, Li Hui,Zhu Zhong-liang 2010, 14 (40):  7588-7592.  doi: 10.3969/j.issn.1673-8225.2010.40.041 Abstract ( 311 )   PDF (303KB) ( 393 )   Save BACKGROUND: Electromagnetic fields can cause changes of the body, especially the nervous system. Effect of pulsed electromagnetic fields (PEMFs) on neural stem cells has been detected. OBJECTIVE: To investigate the effect of prenatal pulsed electromagnetic fields (PPEMFs) on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspring. METHODS: Sprague Dawley female rats weighing 240-260 g were included and randomly divided into two groups: control and PPEMFs. Rats from the control group were given no interventions. Rats from the PPEMFs group were given PEMFs stress at gestational days 14-20. Each stress was given three times daily for 10 minutes. The male and female offspring rats were sacrificed at 1 month of age and their brains were sectioned to determine the expression of nestin protein and Brdu-positive cells in the hippocampus by immunohistochemistry. RESULTS: The expression of nestin- and Brdu-positive cells in the hippocampus of female and male PEMFS offspring were significantly higher compared with the control group (P < 0.001), and there was a significant difference between female and male offspring (P < 0.001). The nestin- and Brdu-positive cells in female offspring outnumbered those in male offspring (P < 0.001); however, there was no significant difference between female and male offspring in the control group (P > 0.05). CONCLUSION: PPEMFs can increase the number and proliferative capability of the neural stem cells in offspring. It may be a primary stage of the cascade reaction of the body to the brain damage caused by PPEMFs stress. Related Articles | Metrics

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Preparation of canine bone marrow stromal stem cell sheets and investigation on their osteoblastic differentiation Bu Ling-xue, Jing Heng, Chen Li-qiang, Gao Zhen-hua, Li Ning-yi 2010, 14 (40):  7593-7596.  doi: 10.3969/j.issn.1673-8225.2010.40.042 Abstract ( 295 )   PDF (285KB) ( 385 )   Save BACKGROUND: Conventional methods, including trypsin digestion and cells transfer using single cell suspension, have many drawbacks, which limit the development of bone tissue engineering. OBJECTIVE: To culture bone marrow stromal stem cells, induce osteoblastic differentiation, and prepare cell sheets. METHODS: Canine bone marrow stromal cells were isolated by density gradient centrifugation technique, inoculated into DMEM medium, and induced to differentiate into osteoblasts. Complete cell sheets were harvested by cell sheet engineering based on the temperature change of temperature-responsive medium. RESULTS AND CONCLUSION: Immediately after inoculation, primary cells were scattered on the bottom of culture flask, presenting a transparent spherical body with a good refractive capacity. At 12 hours, cells exhibited a long shuttle shape, reached complete confluency, and grew in a whirlpool-like fashion. After osteoblastic induction, the majority of bone marrow stromal stem cells appeared tetragonal, polygonal, and squamose. At 21-28 days, round or oval-shaped calcified nodules formed. When the bone marrow stromal stem cells in the temperature-responsive culture dishes were cooled below the critical temperature 32 ℃, cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem cell sheets. These findings indicate that density gradient centrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and cell sheet engineering enables to harvest complete bone marrow stromal stem cell sheets.  Related Articles | Metrics

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Myogenic differentiation factors and 5-azacytidine induce the differentiation of rat bone marrow mesenchymal stem cells into skeletal muscle cells in vitro　 Chen Zhen-qiang, Sun Zhan-sheng, Zhi Wei 2010, 14 (40):  7597-7600.  doi: 10.3969/j.issn.1673-8225.2010.40.043 Abstract ( 298 )   PDF (285KB) ( 314 )   Save BACKGROUND: Muscle transposition is a conventional method to treat muscle tissue defects, but it results in damage to another piece of muscle. For this reason, we designed this study to search for a method to in situ repair muscle tissue defects. OBJECTIVE: To investigate the conditions for in vitro induced differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into skeletal muscle cells. METHODS: Following isolation and culture, passage 3 BMSCs were induced to differentiate in vitro by a combination of 5-azacytidine, myogenic differentiation factor, transforming growth factor β1, and insulin like growth factor. At 9 days after induction, cells were harvested and identified by immunohistochemistry. RESULTS AND CONCLUSION: Primary cultured BMSCs exhibited an adherent, colony-like growth. After 5-7 days, multi-synaptic cells, thin and flat polygonal cells, polygonal cells, and triangle-shaped cells were observed. After 12 days, cells confluenced and covered the whole bottom of culture flask, with slightly altered morphology of BMSCs. After 5-azacytidine induction, some cells died and grew slowly. After 7 days, cells markedly grew and soma was gradually enlarged, presenting with an oval, spindle-shaped, or irregular appearance. After 14 days, spindle-shaped cells become more. After 18-22 days, myotubes were increased in number and enlarged in volume, and myotube nucleuses were also increased. The newly formed myotubes and spindle-shaped fibroblasts were distributed in parallel interval. The immunohistochemistry of BMSCs revealed that cells were positive for CD44, with dark brown granules in the cytoplasm, especially around the nucleus, but they were negative for CD34. The immunohistochemistry of induced BMSCs demonstrated that cells were positive for desmin and skeletal muscle myosins. These findings indicate that myogenic differentiation factors and 5-azacytidine could induce the oriented differentiation of BMSCs into skeletal cells, with the presence of positive expression of desmin and skeletal muscle myosins. Related Articles | Metrics