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04 June 2010, Volume 14 Issue 23 Previous Issue    Next Issue

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Effects of human myocardial extracts on transdifferentiation of bone marrow mesenchymal stem cells and temporal expression of related genes Zeng Jun-yi, Wei Yun-feng, Wang Yang, Xie An, Lou Yuan-lei, Zhang Mei 2010, 14 (23):  4181-4185.  doi: 10.3969/j.issn.1673-8225.2010.23.001 Abstract ( 279 )   PDF (488KB) ( 424 )   Save BACKGROUND: Many studies on animals have suggested that bone marrow mesenchymal stem cells (BMSCs) have the potential to transdifferentiate into cardiomyocytes (CMs) within the myocardial environment both in vivo and in vitro. However, few studies in humans are reported in this field and the molecular signals that underlie this process are not fully understood. OBJECTIVE: To investigate the effects of human myocardial extracts on transdifferentiation of BMSCs and temporal expression of related genes. METHODS: Human BMSCs were harvested in vitro. Human BMSCs from passage 4 were induced by high level of autologous myocardial extracts for 2 weeks. Cardiomyogenic differentiation was confirmed by phase contrast microscope and immunostaining against α-actin and cardiac troponin-T (cTnT), and reverse transcriptase-polymerase chain reaction (RT-PCR) was performed for temporal expression of related genes Nkx-2.5, GATA-4, MEF-2C, α-MHC and cTnT. RESULTS AND CONCLUSION: After induced by myocardial extracts, BMSCs showed a stick-like morphology, with a greater volume and stronger three-dimensional sense. 25.53% of BMSCs were positive for α-actin and 23.48% for cTnT at 2 weeks. The expression level of Nkx-2.5, GATA-4 and MEF-2C genes began to increase at 3 days, the expression level of GATA-4 and MEF-2C genes reached a peak at 5 days after induction and remained at relatively high level afterwards, and the expression level of Nkx-2.5 was gradually increased with the induction time. The expression of α-MHC and cTnT genes, which were not detected before induction, began to emerge at 3 days after induction and remained at high level afterwards. Therefore, myocardial extracts are conducive for BMSCs to differentiate into CMs in vitro, along with the temporal expression changes of Nkx-2.5, GATA-4, MEF-2C, α-MHC and cTnT genes during the transdifferentiation. Related Articles | Metrics

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Construction and expression of an adenoviral vector encoding human interleukin-12 in human bone marrow mesenchymal stem cells Chen Yuan-yuan, Tan Xiao-hua, Ma Jing-ying 2010, 14 (23):  4186-4190.  doi: 10.3969/j.issn.1673-8225.2010.23.002 Abstract ( 258 )   PDF (463KB) ( 500 )   Save BACKGROUND: Previous studies have suggested that interleukin-12 (IL-12) is a powerfull anti-tumor cell factor. Bone marrow mesenchymal stem cells (BMSCs) are prone to introduction and expression of exogenous gene, and have weak immunizing antigen and immunological regulation functions. Therefore, mesenchymal stem cells (MSCs) carrying IL-12 gene possess great prospects for tumor treatment. OBJECTIVE: To construct an adenoviral (Ad) vector encoding human IL-12 (hIL-12) gene, infect human BMSCs, and detect the expression of IL-12. METHODS: The total RNA of human dendritic cells (DCs) was obtained. hIL-12 p35 and p40 cDNA were amplified from the total RNA by RT-PCR, and p35 and p40 fragments were linked by internal ribosomal entry sites (IRES), to construct the shuttle vector pDC515 p35/IRES/p40. Using Ad MaxTM adenovirus vector system, pDC515 p35/IRES/p40 and pBHGfrt△E1, 3FLP were cotransfected into 293 cells, and Ad hIL-12 was generated by FLP recombinase-mediated site-specific recombination. After Ad hIL-12 infection, hBMSCs were irradiated with γ-ray to lose proliferative activity. IL-12 levels were determined in cell supernatants utilizing hIL12p70 enzyme linked immunosorbent assay. RESULTS AND CONCLUSION: The sequences of p35 and p40 fragments were identical with those provided by GenBank NM_000882 (762 bp) and NM_002187 (987 bp), respectively. Ad MaxTM was an efficiently and quickly packaging system of adenoviral vectors. hIL-12 gene in which the two subunits p35 and p40 were linked by IRES can efficiently express the protein of IL-12p70. The high expression of IL-12 is consecutively found after the infection of human BMSCs with Ad hIL-12, suggesting a potential application to the gene therapy of human BMSCs as a carrier. Related Articles | Metrics

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Simvastatin effects on correlation factor expression of Wnt signaling pathway in rat bone marrow stromal stem cells Zheng Huan, Zhang Liu, Tian Fa-ming, Zhang Hui, Mu Shu-lin 2010, 14 (23):  4191-4194.  doi: 10.3969/j.issn.1673-8225.2010.23.003 Abstract ( 445 )   PDF (426KB) ( 342 )   Save BACKGROUND: As a lipid-lowering drug, simvastatin is getting more and more attention for its potential effects on bone formation in research of bone metabolism, but contradiction still exists when it is used for in vivo study. OBJECTIVE: To investigate the effects of simvastatin on bone mass, proliferation and differentiation of the cultured bone marrow stromal cells (BMSCs) in rats, as well as mRNA expression levels of some related factors of Wnt signaling pathway. METHODS: A total of 36 6-week-old female Sprague-Dawley rats were randomized into two groups. In the control group, the rats were treated with distilled water daily by gavage. In the experimental group, the rats were administrated 20 mg/kg simvastatin per day. Six rats from either group were sacrificed after the final administration at 3, 6, 9 weeks separately. The left femora were removed for the measurement of bone mineral density (BMD). BMSCs derived from the right femora and tibiae were cultured in osteoblastic differentiation-induced medium. Alkaline phosphatase (ALP) activity measurement and ALP staining were performed on day 16. Real-time PCR was used to evaluate the mRNA expression levels of LRP-5, Axin2 and β-catenin on day 21 following total RNA was extracted. RESULTS AND CONCLUSION: After being administrated with simvastatin for 3, 6 and 9 weeks, BMD of rats had no significant change. There were no significant differences in gene mRNA levels and osteogenic differentiation potential in BMSCs cultured in vitro compared with the control group. Administrated with simvastatin for either 3, 6 or 9 weeks had no significant effect on BMD and the differentiation of BMSCs in rats, and on the expression levels of LRP-5, Axin2 and β-catenin mRNA. Related Articles | Metrics

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In vitro proliferation and multi-directional differentiation of rat bone marrow mesenchymal stem cells Xu Yong-tao, Li Feng, Lu Hou-geng 2010, 14 (23):  4195-4198.  doi: 10.3969/j.issn.1673-8225.2010.23.004 Abstract ( 299 )   PDF (581KB) ( 378 )   Save BACKGROUND: Proliferation of bone marrow mesenchymal stem cells is not clear under some physiological condition; however, mitosis occurs following stimulation to a certain degree. The bone marrow mesenchymal stem cells have a strong proliferation ability and multi-directional differentiation potency. OBJECTIVE: To further investigate the in vitro proliferation and multi-directional differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were separated using whole bone marrow culture method and purified using attachment method. Morphology and growth characteristics were observed under inverted phase contrast microscope and growth curve was drawn using MTT method. Immunohistochemical method was used to identify cell surface markers CD44 of stem cells. The fourth-passage cells were respectively cultured in osteogenic, adipogenic and neuro-differentiation induction medium, and confirmed using the alkaline phosphatase staining, type II collagen immunohistochemical staining, oil red O staining and NeuN antibody immunohistochemical staining. RESULTS AND CONCLUSION: After isolation and culture, cells developed a spindle or polygonal appearance. Growth curve was S-shaped, and population doubling time was 31 hours. Immunohistochemical staining demonstrated that CD44 expression was positive, but CD34 was negative. Two weeks after induction, calcium salt mineralization was found in the bone marrow mesenchymal stem cells of passage 4. Two weeks after chondrogenic induction, type II collagen demonstrated a positive expression. Two weeks after fat-induced culture, a great quantity of red fat drops were observed in cytoplasm. Six hours after neural induction, cells displayed processes, which were similar to axon and dendrite fiber. NeuN immunohistochemical staining demonstrated a positive result. This suggested that in vitro cultured bone marrow mesenchymal stem cells had a strong proliferation and could differentiate into osteoblasts, chondrocytes, adipocytes, and neuronal-like cells. Related Articles | Metrics

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Effects of liposome-mediated human bone morphogenetic protein-2 gene transfection on growth and proliferation of rabbit adipose-derived stem cells An Rong-ze, Liu Fan-fan, Wang Zhao-jie, Yuan Xiao-hong, Qi Xin-wen, Zhao Jun-yan, Yang Jin, Zhao Hao, Hu Xiao-jun,Chen Jun-ping, Wen Guang-yu 2010, 14 (23):  4199-4202.  doi: 10.3969/j.issn.1673-8225.2010.23.005 Abstract ( 346 )   Save BACKGROUND: Liposome has toxic side effect on cells, but introduction of exogenous genes into cells affects cell metabolism. Therefore, it is worthy to explore effects of liposome-mediated human bone morphogenetic protein-2 (hBMP-2) gene-transfected adipose-derived stem cells (ADSCs) on cell growth and proliferation. OBJECTIVE: To investigate the effects of liposome-mediated hBMP-2 gene transfection on growth and proliferation of rabbit ADSCs. METHODS: The rabbit ADSCs were extracted from the adipose tissue and subcultured in vitro. The fourth passage of ADSCs were collected and transfected by hBMP-2 gene and enhanced green fluorescent protein (EGFP) gene respectively. The cell morphology and their related growth pattern were continuously observed under the inverted microscope after transfection. At 48 hours after transfection, the transient transfection efficiency was measured by fluorescence microscopic counting. MTT assay was used to drawn the cell growth curve and to calculate the colony doubling time. RESULTS AND CONCLUSION: At 48 hours after transfection, cell volume became large, with abundant cytoplasm, and some were round or oval. The transient transfection rate was about (18.0±0.42)% under fluorescence microscope. According to the cell growth curves drawn by MTT assay, the transfected cells grow at a slower rate compared with the untransfected cells, but both of them roughly had the same growth curve, like the shape of “S”, and little change in their overall trends. After transfection, the population doubling time was 55.51 hours in the hBMP-2 group, 53.58 hours in the EGFP group and 46.10 hours in the non-transfected group. There was no significant statistical difference (P > 0.05). Results have indicated that liposomes can mediate the hBMP-2 gene and EGFP gene transfected rabbit ADSCs and it has no effect on their growth and proliferation. Related Articles | Metrics

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Osteogenic activity of in vitro cultured human adipose stromal cells versus bone marrow stromal cells Jiang Yi, Liu Zhi, Han Li-qiang, Zhu Zheng-yan, Tian Yong-gang 2010, 14 (23):  4203-4206.  doi: 10.3969/j.issn.1673-8225.2010.23.006 Abstract ( 296 )   PDF (431KB) ( 547 )   Save BACKGROUND: In present research of tissue engineered bone construction, seed cells are mainly harvested from bone, periosteum, bone marrow and non-osseous tissue. Present studies mainly focused on bone marrow stromal cells (BMSCs), but the discovery of adipose-derived stromal cells (ADSCs) can replace bone marrow stromal cells. OBJECTIVE: To study the biological characteristic of ADSCs and BMSCs cultured in vitro and to compare alkaline phosphatase activities following osteogenesis so as to evaluate whether ADSCs could be a better seed cells to replace BMSCs. METHODS: Adipose tissue and bone marrow from the same body were collected during operation. Adipose tissues were digested with collagenase type Ⅰ to gain ADSCs, and BMSCs were isolated by centrifugation on lymphocyte separation medium. Following in vitro culture and passage, cells were induced to differentiate into osteoblasts. At the second and third weeks after induction, alkaline phosphatase activity in cells was measured for comparison and Von Kossa calcium nodus staining was used to detect osteoblasts. RESULTS AND CONCLUSION: Adipose tissue and bone marrow were gained from 15 patients and 10 of them brought off a successful experiment. Compared with BMSCs, ADSCs can be cultured more easily and proliferate faster. Their morphological characteristics were similar. Black calcium noduses were seen in extracellular matrix. No significant difference was determined in alkaline phosphatase activity between ADSCs and BMSCs (P > 0.05). The results proved that adipose tissue is abundant. ADSCs can be cultured more easily, and have similar biological characteristics with BMSCs, and they can be easily cultured, proliferate faster, and can be induced to osteoblasts. ADSCs should be a more ideal seed cells than BMSCs Related Articles | Metrics

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Immunophenotype determination of rat adipose tissue-derived mesenchymal stem cells at passage 4 Li Dong-fei, Yang Chun, Li Zhen, Dai Jing-xing, Yuan Lin 2010, 14 (23):  4207-4210.  doi: 10.3969/j.issn.1673-8225.2010.23.007 Abstract ( 331 )   PDF (470KB) ( 436 )   Save BACKGROUND: Many studies have confirmed that adipose tissue-derived mesenchymal stem cells (ADMSCs) can be collected using enzyme digestion method. At passage 10, ADMSCs grew well, but their surface marker expression decreases. OBJECTIVE: To isolate and cultivate rat ADMSCs and detect immunophenotype at passage 4. METHODS: ADMSCs of six Sprague-Dawley rats from inguinal fat pads were isolated and cultured. Adherent cells were amplified till passage 4. Morphology of ADMSCs was observed under a phase contrast microscope. The immunophenotype markers including CD11b, CD29, CD45, CD49d, CD90 and CD106 of ADMSCs were determined using flow cytometry.  RESULTS AND CONCLUSION: Rat ADMSCs showed fibroblastic-like morphologic characteristics. The immunophenotype analysis revealed that the rat ADMSCs were positive for CD29 and CD90, but negative for CD11b, CD45, CD49d and CD106. The rat ADMSCs can be obtained through enzyme-based isolation procedures. The results revealed that the immunophenotype characterization of rat ADMSCs is consistent with mesenchymal stem cells. Related Articles | Metrics

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Biological characteristics of mesenchymal stem cells from bone marrow versus adipose tissue in rats Yang Zhong-meng, Yin Zhen-yu, Ye Zhi-jian, Zhang Qiong, Zhao Wen-xiu, Wang Xiao-min 2010, 14 (23):  4211-4216.  doi: 10.3969/j.issn.1673-8225.2010.23.008 Abstract ( 348 )   PDF (667KB) ( 483 )   Save BACKGROUND: Except bone marrow, people have collected mesenchymal stem cells (MSCs) from placenta tissue, cord blood muscle tissue and adipose tissue. OBJECTIVE: To compare the difference in biological characteristics and immunoloregulation of rat adipose-derived MSCs (ADSCs) and bone marrow MSCs (BMSCs). METHODS: ADSCs and BMSCs were obtained respectively from adipose and bone marrow of BN rats. ADSCs and BMSCs were isolated and purified in vitro to determine cell morphology, surface marker, growth kinetics and differentiation potential. Immunoloregulation of ADSCs and BMSCs was compared in mixed lymphocyte response RESULTS AND CONCLUSION: Morphology was similar between ADSCs and BMSCs under an optical microscope and transmission electron microscope. ADSCs and BMSCs at passage 3 highly expressed CD29 and CD90, weakly expressed CD34, CD45 and CD11b. The proliferation speed of ADSCs at passages 3, 4 and 5 was obviously faster than BMSCs. Both ADSCs and BMSCs possessed low immunogenicity, could suppress allogenic antigen-induced T lymphocyte proliferation. Moreover, this inhibitory effect was positively associated with cell number. There were no significant differences in inhibitory effects of an equal volume of ADSCs and BMSCs. Results verified that ADSCs had the same low immunogenicity and immunoloregulation as BMSCs. Related Articles | Metrics

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Transforming growth factor-beta 1/ insulin-like growth factor-1 versus nucleus pulposus cells in inducing the differentiation of adipose-derived mesenchymal stem cells into nucleus pulposus-like cells Liu Lian-jiang, Li Fang, Wen Tian-yong 2010, 14 (23):  4217-4221.  doi: 10.3969/j.issn.1673-8225.2010.23.009 Abstract ( 316 )   PDF (521KB) ( 385 )   Save BACKGROUND: Previous studies have addressed adipose-derived mesenchymal stem cells (ADSCs) culture way for monolayer culture; induction way was mainly cytokine method, and under intervertebral discs micro-environment, cell interactions also deserved further study in its three-dimensional environment, in addition to the effects on cell cytokines. OBJECTIVE: To observe differences in differentiation of ADSCs into NPCs under two induced conditions: nucleus pulposus cells (NPCs) and transforming growth factor β1 (TGF-β1)/ insulin-like growth factor (IGF-1) in vitro. METHODS: Rabbit ADSCs monolayer culture and NPCs were cultured respectively, 3 generations of ADSCs were qualified; 5×106 cells were made into microspheres and placed in Transwell culture plates for culture under TGF-β 1/IGF-1 induction. Cell shape changes were observed before, 7, 14 days after induction. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine levels of type II collagen and proteoglycan. RESULTS AND CONCLUSION: At 7 and 14 days, no significant difference was detected in size and shape of ADSCs microspheres in both groups. RT-PCR detection results displayed that type II collagen and proteoglycan mRNA expression was determined in 7-day TGF-β1/IGF-1 group and NPCs-induced group, but expression was stronger in the TGF-β1/IGF-1 group. Type II collagen and proteoglycan mRNA expression was obviously increased in 14-day NPCs-induced group, superior to TGF-β1/IGF-1 group. Results have suggested that in vitro NPCs and TGF-β1/IGF-1 lead to the ADSCs, which had promoting effects on NPCs differentiation. TGF-β 1 and IGF-1 are normal NPCs-secreted important factors. The promoting effects on proliferation and differentiation exist, with the exception of cytokine induction effect, in differentiation of ADSCs into NPCs. Related Articles | Metrics

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Effect of microglia on differentiation from in vitro cultured neural stem cell into cholinergic neurons Jin Yu-ling, Liu Qie, Liu Jing-jing, Li Xin, Wang Li-hua 2010, 14 (23):  4222-4226.  doi: 10.3969/j.issn.1673-8225.2010.23.010 Abstract ( 302 )   PDF (554KB) ( 510 )   Save BACKGROUND: Orient differentiation of neural stem cells depends on concentration of microglia. A small concentration can not play an effect to a certain degree; and a large concentration might cause cytotoxicity. OBJECTIVE: To observe the effect of varying concentrations of microglia on differentiation from in vitro cultured neural stem cells into neurons. METHODS: Newborn Wistar rats within 24 and 48 hours were used for primary culture of neural stem cells and microglia, which were then identified by immunohistochemistry. Neural stem cells and microglia were co-inoculated in 6-well plates, at the density of 10:1, 4:1, 1:1, 1:4, and 1:10, with 6 wells at each density. Neural stem cells alone were considered as control group and incubated for days 3, 7 and 14 to observe the cell growth. RESULTS AND CONCLUSION: Primary culture of neural stem cells gathered like a ball and grew in a suspending manner. Nestin staining was positive, but nuclei were not stained. Microglia grew in an adherent manner, with strong refractive index. Most of the cells showed short branch-like protrusions. Microglia-specific marker antibodies CD11b/c staining showed that the cell purity was above 80%. Compared with control group, neural stem cells and microglia cells were co-cultured at a density of 10:1, 4:1, 1:1, 1:4, and 1:1. ChAT-positive cells were significantly increased. If neural stem cells and microglia were co-cultured at the density of 4:1, the increasing was significantly greater than other co-cultured statuses (P < 0.05), and the cell growth peaked at 7 days. The microglia could promote differentiation from neural stem cells into cholinergic neurons, and the co-culture of neural stem cells and microglia at a density of 4:1 achieved the best effect at 7 days. Related Articles | Metrics

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Induction and differentiation of bone marrow mesenchymal stem cells into dopaminergic neurons Liu Zhuo, Xu Yun, Huang Dan-qing 2010, 14 (23):  4227-4230.  doi: 10.3969/j.issn.1673-8225.2010.23.011 Abstract ( 448 )   PDF (414KB) ( 368 )   Save BACKGROUND: Recent studies have found that neurotrophic factor plays important roles in differentiation of bone mesenychymal stem cells (BMSCs). However, few reports have addressed whether neural stem cells (NSCs) with regeneration capacity in the brain tissue in vitro can induce the differentiation of BMSCs into dopaminergic neurons. OBJECTIVE: To investigate the possibility of rat BMSCs differentiation into dopaminergic neurons under the induction condition of glial cells-derived neurotrophic factor (GDNF) and NSCs coculture. METHODS: BMSCs from Sprague Dawley rats were isolated and cultured. The third passage of BMSCs was divided into two groups. Group A: Medium was MEM containing GDNF; group B: BMSCs marked with Brdu and then cocultured with NSCs which had formed primary neurospheres. Three days later, BMSCs from each group were suffered immunohistochemistry staining to examine the tyrosine hydroxylase of the differentiated neuron to observe the differentiation of BMSCs. RESULTS AND CONCLUSION: At 24 hours postinduction, BMSCs of group A were cone-shaped and had the prolonged processes. The amount of processes increased, with the presence of neuron-like morphology. The web connection among the cells can also be observed. Three days later, some cells expressed tyrosine hydroxylase. In group B, NSCs attached to culture dish immediately. The attached cells proliferated and showed neuronal phenotype. A majority of attached cells expressed tyrosine hydroxylase, and a few cells presented double staining: Brdu/tyrosine hydroxylase. Results have exhibited that BMSCs have the potential of differentiating into dopaminergic neurons. The GDNF proves to be more effective in the experiment compared with NSCs. Related Articles | Metrics

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Distribution of bone marrow mesenchymal stem cells intravenously transplantated in chronic myocardial infarction rats Yang Zhi-kai, Wang Wei, Gao Bing-ren, Jin Pei-feng, Hu Sheng-shou, Zhang Hao 2010, 14 (23):  4231-4234.  doi: 10.3969/j.issn.1673-8225.2010.23.012 Abstract ( 453 )   PDF (379KB) ( 360 )   Save BACKGROUND: Present studies mainly focused on acute myocardial infarction in research of cell transplantation. There are a few studies addressing stem cells intravenously transplanted following chronic myocardial infarction. In particular, there are no reports concerning distribution and survival of transplanted stem cells at various time points following transplantation by dynamic monitoring. OBJECTIVE: To observe distribution of bone marrow mesenchymal stem cells (BMSCs) intravenously transplanted in rats with chronic myocardial infarction. METHODS: BMSCs from male Sprague Dawley rats were cultured for transplantation. Three weeks after induction of myocardial infarction, 18 female Sprague Dawley rats were randomized into 2 groups. Suspension of phosphate buffered saline (PBS) and BMSCs (5×106, 300 μL) were injected into the femoral vein of experimental group (n = 9). Meanwhile, an equal volume of PBS was injected into the femoral vein of control group (n = 9). Another 9 female Sprague Dawley rats that underwent thoracotomy were only involved as sham operation group, without ligation of coronary artery. Suspension containing BMSCs (5×106, 300 μL) was injected by the same way in the sham operation group. 300 μL suspension supplemented with 5×106 BMSCs was infused in the femoral vein. At 24 hours, 2 weeks and 1 month following BMSCs transplantation, BMSCs distribution was evaluated through real-time fluorescent quantitative polymerase chain reaction. RESULTS AND CONCLUSION: At 1 day after BMSCs injection, no significant difference in cell distribution was detected in the lung, liver and spleen (P > 0.05). However, cell number in the heart was significantly greater in the experimental group than in the sham operation group (P < 0.01). At 2 weeks following transplantation, cell number was significantly decreased in the lung in both groups (P < 0.05), and there were no significant differences in cell distribution among liver, spleen and heart (P > 0.05). At 4 weeks following transplantation, no significant difference was detected in the lung, liver and spleen in both groups (P > 0.05). No cells were detected in the heart in the sham operation group. At 4 weeks following transplantation, cell distribution was significantly decreased in various tissues. SRY gene was not detected at various time points in the control group. Above-mentioned results have suggested that a large number of BMSCs intravenously transplanted were detected in the lung in rats with chronic myocardial infarction. Moreover, cell number was decreased over time. In addition, a few cells were detected in the heart, spleen and liver. Related Articles | Metrics

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Tacrolimus promotes repair and regeneration of injured spinal cord following neural stem cells transplantation in rats Wu Qiao-li, Li Qing-guo, Huang Hui-ling, Liang Jian-wei, Cao Yi-bo, Sun Yi-lin, Liu Kui 2010, 14 (23):  4235-4238.  doi: 10.3969/j.issn.1673-8225.2010.23.013 Abstract ( 299 )   PDF (550KB) ( 450 )   Save BACKGROUND: Studies have confirmed that tacrolimus not only inhibits T cell proliferation and activation, but also suppresses microglia, macrophages and other inflammatory cells to aggregate, activate and release associated inflammatory cytokines in injuries, which can reduce the damage of secondary inflammation to surrounding normal tissue, thus, provide protective effect to nerves in injuries.  OBJECTIVE: To observe the effect of tacrolimus on spinal cord regeneration and injury repair following neural stem cells (NSCs) transplantation in rats. METHODS: NSCs were isolated form 13-day-pregnant SD rats. Aneurysm clipping folder was used to clip T8 spinal cord of rats under microscope to establish animal model of spinal cord compression injury. At 7 days after injury, all rats were randomly divided into 3 groups: rats in the control group were injected with normal saline at the injury center orientation; those in the cell transplantation group received center directional injection of neural stem cells (NSCs); those in the tacrolimus (FK506) group received directional injection of NSCs combined with intraperitoneal injection of immunosuppressant FK506, 1 mm/kg per day for 7 continuous days. The spinal cord tissue regeneration and neuronal changes were observed using BDA anterograde tracer, haematoxylin-eosin staining, immunohistochemistry and electron microscopy at 1, 2, 4 and 8 weeks after operation. RESULTS AND CONCLUSION: There were no fibers passing through the distal end of the injury center in the control group. However, some nerve fibers in the cell transplantation and FK506 groups passed at 1 week after treatment. Parts of regenerating BDA-positive corticospinal tract passed through the site of spinal cord injury in the cell transplantation and FK506 groups at 8 weeks after treatment, especially in the FK506 group, which may be continued to 1.7 cm of the injury center. Hematoxylin-eosin staining showed that, the necrotic foci began to shrink, and the foam cells decreased in the cell transplantation group and FK506 group at 2 weeks after treatment. Electron microscopy results showed that, in the FK506 group, more normal microfilament and microtubule structure appeared at 1 week, the astrocytes, Schwann cells, and myelin sheaths were commonly seen at 8 weeks after treatment, nerve axons of excitatory terminals had more excitatory transmitter and non-typical tree-axis connections, and there were more normal myelin sheaths. It illustrated that, NSCs transplantation combined with FK506 can reduce the early post-acute inflammatory response, ensure the survival of nerve cells, exhibit neuroprotective and neurotrophic roles, as well as speed up the recovery of neurological function. Related Articles | Metrics

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Striatal dopamine content in Parkinson’s disease rats following human umbilical cord blood mesenchymal stem cell transplantation Liu Fang, Fan Zhi-gang, Han Xue-fei 2010, 14 (23):  4239-4242.  doi: 10.3969/j.issn.1673-8225.2010.23.014 Abstract ( 359 )   PDF (467KB) ( 581 )   Save BACKGROUND: At present, researches of stem cell transplantation for Parkinson’s disease (PD) animal model mainly utilize bone marrow mesenchymal stem cells (MSCs) and embryonic stem cells. Researches of umbilical cord blood MSCs transplantation are relatively less, and there is no report regarding the change of striatal dopamine content before and after umbilical cord blood MSC transplantation. OBJECTIVE: To explore the effect of human umbilical cord blood MSCs transplantation on the dopamine content in PD rat striatum. METHODS: The PD rat models were randomly divided into experimental (n=20) and control (n=20) groups. The third passage MSCs were marked by Hoechst33258, and stereotactically transplanted into experimental group, and the control group was given PBS. The brain tissue of PD rat was sectioned to identify the survival and migration of MSCs and the expression of neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP), tyrosine hydroxylase (TH), and synaptophysin under fluorescent microscope at weeks 2, 4 and 8. Dopamine in striatum was detected by high performance liquid chromatography at 8 weeks of transplantation. RESULTS AND CONCLUSION: Transplanted MSCs survived in the brain, distributed in striatum, callus, and cortex with increasing time, and expressed NSE, GFAP, and TH, but not synapsin. The content of dopamine was significantly increased than control group (P < 0. 05). Related Articles | Metrics

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Migration and localization of bone marrow mesenchymal stem cells labeled by DAPI in the brain of glioma rat models Yu Yin, Yang Hong-fa, Shao Jia-jia, Wu Wei, Zhu Dong, Jiang Tao, Guo Yong-chuan, Liang Qian-lei, Li Yan-xin 2010, 14 (23):  4243-4246.  doi: 10.3969/j.issn.1673-8225.2010.23.015 Abstract ( 348 )   PDF (418KB) ( 587 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) as vector cells were transplanted in the brain for treating glioma. How to verify BMSCs are best drug carriers. OBJECTIVE: To observe the migration and localization of BMSCs in tumor following transplantation of rat BMSCs labeled by DAPI in the brain of a rat model. METHODS: BMSCs were cultured in vitro. Using stereotaxic apparatus, C6 cells were injected into rat brain to establish glioma models. BMSCs labeled by DAPI were injected into the rat brain using a microinjector. The rats were sacrificed at 3 and 7 days following transplantation. The localization of BMSCs in tumor and the relationship with tumor cells were observed under a confocal microscope. RESULTS AND CONCLUSION: The glioma model in rat brain was established successfully. BMSCs labeled by DAPI can accumulate around tumor vessel actively in rat brain, and can mix together with tumor cell. These suggested that BMSCs are a good carrier of gene therapy. Related Articles | Metrics

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Effects of 5-azacytidine-induced bone marrow mesenchymal stem cells transplantation on cardiac function Chen Li-xing, Nie Jun, Sun Jin-hua, Cai Hong-yan, Zhao Ling, Guo Tao, Xiao Jian-ming 2010, 14 (23):  4247-4251.  doi: 10.3969/j.issn.1673-8225.2010.23.016 Abstract ( 253 )   PDF (453KB) ( 411 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into various cells. BMSCs induced by 5-azacytidine were transplanted into myocardium of rabbits after myocardial infarction. Whether transplanted stem cells could improve cardiac function in short or long time was unclear. OBJECTIVE: To investigate the effects of BMSCs transplantation on cardiac function in rabbits after acute myocardial infarction (AMI). METHODS: Rabbit BMSCs were purified and cultured in vitro. BMSCs were induced by 5-azacytidine, and labeled with DAPI at 4 weeks following induction. New Zealand rabbits were randomly divided into three groups. In sham operation group, the chest was opened for 1 hour and then closed. In BMSCs group, autologous BMSCs were infused into the surrounding infarct area at 4 points using a microsyringe at 1 hour following model induction by coronary artery ligation. In AMI group, an equal volume of saline was injected at the same region following model establishment. At 3 days and 4 weeks following transplantation, ultrasonic cardiography was used to determine changes in left ventricular end diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and left ventricular ejection fraction (LVEF) in rats from each group. RESULTS AND CONCLUSION: Fluorescence microscopy results demonstrated that DAPI-labeled BMSCs were distributed extensively in the myocardium of BMSCs group at 3 days and 4 weeks, arranged in parallel with the cardiac muscle fibers. At 3 days after transplantation, there were no significant differences in LVEDV, LVESV and LVEF between AMI and BMSCs groups. No significant improvement was found in cardiac function. At 4 weeks after transplantation, LVEDV and LVESV were significantly decreased, and LVEF was significantly increased in BMSCs group compared with AMI group. Related Articles | Metrics

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Bone marrow mesenchymal stem cells in repair of injured pancreas in rats Chen Qiang, Wang Qiu-jing, Yang Dong-xu, Zhou Yu-lai 2010, 14 (23):  4252-4256.  doi: 10.3969/j.issn.1673-8225.2010.23.017 Abstract ( 269 )   PDF (497KB) ( 495 )   Save BACKGROUND: The ischemic injury of pancreas is a common practical problem in the clinic, which is difficult to solve. The routine methods of prevention and treatment get little success. Bone marrow mesenchymal stem cells (BMSCs) are a widely accepted modus that is developed in the recent years to treat injury, so it is necessary to find an effective way to treat the ischemic injury of pancreas using stem cells. OBJECTIVE: To study the repair effect of BMSCs on ischemic injury of pancreas. METHODS: Adherent culture and tissue culture were used to get highly purified BMSCs, which were incubated in L-DMEM containing 20% fetal bovine serum. Following three subcultures, BMSCs were labeled by enhanced green fluorescent protein recombinant adenovirus by injection into local pancreas of animal models where ischemic injury happened. Survival rate of the rats was observed, and the position of the marked cells in frozen sections was observed by confocal microscopy. RESULTS AND CONCLUSION: Highly purified BMSCs were harvested from adherent culture. The survival rate was significantly higher in injected animals than the control rats. The results of confocal microscopy indicated that after 24 hours of the injection, the cells reached the ischemic part and aggregated in the microcirculatory system, and then entered the injured part gradually and heavily, finally got highly expressed in the microcirculatory system and conduit system. Results have suggested that the survival rate of the rats can be heightened by local injection of BMSCs, and this is propitious to the recovery of ischemic pancreas. Related Articles | Metrics

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Transplantation of bone marrow mesenchymal stem cells for repair of chronic pancreatic injury in rats Liu Hong-bin, Yang Jing, Li Dong-hua, Wang Qian 2010, 14 (23):  4257-4261.  doi: 10.3969/j.issn.1673-8225.2010.23.018 Abstract ( 265 )   PDF (533KB) ( 511 )   Save BACKGROUND: Chronic pancreatitis is a chronic necroinflammatory process characterized pathologically by chronic inflammation and fibrosis in pancreas tissue. Bone marrow mesenchymal stem cells (BMSCs) maybe play a potential role in treating chronic pancreatitis. However, few reports have addressed the use of MSCs in pancreatic regeneration. OBJECTIVE: To investigate the effects and mechanisms of BMSCs for treating chronic pancreatitis in rats. METHODS: BMSCs were isolated, cultured and identified in vitro. Chronic pancratitis rat model was induced by infusion of oleic acid to bile-pancreas duct in Wistar rats. The Wistar rats were assigned randomly to sham operation group, model group and BMSCs group. BMSCs were transplanted to the rats from BMSCs group through caudal vein injection in the number of 3 ×    106 /mL at 20 days after the model induction and the injection was repeated twice at 40 days and 60 days after model induction. For the model group, the equal volume of saline was injected into the caudal vein. For the sham operation group, duodenum puncture and infusion of oleic acid to bile-pancreas duct were not done. For all the rats, pancreatic tissues were collected and underwent histopathological examination. Pancreatic connective tissue growth factor (CTGF), transforming growth factor β (TGF-β), type I collagen, type III collagen and myeloperoxidase (MPO) activity were detcted. RESULTS AND CONCLUSION: The pathological injury and the fibrosis in the BMSCs group were ameliorated signicantly. The contents of pancreatic CTGF, TGF-β, type I collagen, type III collagen and MPO were all decreased significantly (P < 0.01). These results suggested that BMSCs have obvious repairing effects on the injured pancreatic tissue of chronic pancreatitis rats, which may be related to the inhibition of CTGF, TGF-β release, decreased production of type I, type III collagen, and inhibition of inflammatory reaction. Related Articles | Metrics

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Colonization and differentiation of allogeneic rat bone marrow mesenchymal stem cells transplanted into the intestine with ischemia/reperfusion injury Gao Guang-zhou, Li Da-wei, Li Xin, Sun Tao 2010, 14 (23):  4262-4266.  doi: 10.3969/j.issn.1673-8225.2010.23.019 Abstract ( 345 )   PDF (446KB) ( 484 )   Save BACKGROUND: The small intestine is extremely sensitive to the hypoxia and ischemia, and it would be got severe damage after ischemia/reperfusion injury. Bone marrow mesenchymal stem cells (BMSCs) are multipotent cells which might participate in the repair of damaged tissue through a variety of ways. OBJECTIVE: To investigate colonization and therapeutic effects of allogeneic rat BMSCs transplantation in the intestine of ischemia/reperfusion injury model. METHODS: Wistar female rats were assigned to three groups. The abdominal cavity was opened and then sutured in the sham operation group. Remaining rats were used to establish ischemia/reperfusion models. Rats in the control group were only infused with saline following intestine with ischemia/reperfusion injury. Rats in the treatment group were treated with BMSCs from Wistar male rats via caudal vein. Following treatment, jejunal tissue was harvested at 12, 24 hours, 3, 7, 14 and 28 days separately to make frozen sections. The distribution of donor cells in the receptor intestine was observed under a fluorescence microscope. Reverse transcription-polymerase chain reaction was employed to determine the sex-determining gene (SRY) in male rats. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the jejunal tissues were detected. RESULTS AND CONCLUSION: No donor cell transfused in intestinal villi was detected under the fluorescence microscope. The expression ratio of the SRY gene was 50%, 66.6%, 33.3%, 16.6% at 3, 7, 14, and 28 days respectively. The levels of MDA were lower, but SOD levels were higher in jejunum tissues in the treatment group than the control group on 12, 24 hours, 3 and 7 days. Results have found that BMSCs from allogeneic rats could colonize in the injured intestine of the receptor rat with ischemia/reperfusion injury. The BMSCs transplantation can promote the recovery of intestinal injury. The beneficial effects of BMSCs were primarily mediated via paracrine actions but not by their direct differentiation into target cells. Related Articles | Metrics

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Slow skeletal muscle satellite cells transplantation for repair of skeletal muscle contusion injury Hu Jiu-ting, Zhu Dao-li 2010, 14 (23):  4271-4274.  doi: 10.3969/j.issn.1673-8225.2010.23.021 Abstract ( 392 )   PDF (495KB) ( 407 )   Save BACKGROUND: Muscle satellite cells play an important role in skeletal muscle injury, repair and maintenance. The muscle wet weight, muscle fiber cross-sectional area and the content of actin could provide indicators of the recovery of muscle injury. OBJECTIVE: To investigate the effect of transplantation of slow skeletal muscle satellite cells on repairing muscle contusion damage at 5, 10, 15 days post surgery. METHODS: Slow skeletal muscle satellite cells of Sprague-Dawley rat were cultured in vitro. Slow muscle satellite cells and physiological saline (control group) were respectively implanted into injured position of contusion animal model. The repairable conditions of skeletal muscle injury were observed.  RESULTS AND CONCLUSION: The experimental group post-transplantation obtained good results compared with the control group. ① At 5, 10 and 15 days after the transplantation of satellite cells, satellite cells labeled with fluorescence in muscle tissue gradually reduced in the experimental group. Muscle satellite cells formed myotubes and eventually fused into muscle fibers so as to repair the damage. ②At 5, 10 and 15 days after transplantation, the fibrillation potentials and positive sharp waves were progressively reduced. Myoelectric intensity and pulse width in turn at the injury side were increased by degrees to near normal side, while EMG intensity and pulse width were increased slowly in the control group. ③With the processing of the injured repair, the recovery rates of muscle wet weight were progressively reduced. ④With the processing of the recovery, the gray values of the experimental group were higher than the control group. Results have indicated that the transplantation of slow skeletal muscle satellite cells into the contusion muscles can significantly improve the recovery of muscle contusion damage. Related Articles | Metrics

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Distribution and differentiation of bone marrow mesenchymal stem cells transplanted into vitals using different methods Guo Qi-cang, Wang Yu-fang, Zhang Yu, Wu Peng-yu, Tang Huan-feng, Liu Zhi-yong, Li Zhan-qing 2010, 14 (23):  4275-4280.  doi: 10.3969/j.issn.1673-8225.2010.23.022 Abstract ( 270 )   PDF (628KB) ( 327 )   Save BACKGROUND: Numerous studies have demonstrated that chemotaxis or direct intramyocardial injection of bone marrow stem cells can differentiate into myocardial cells, vascular endothelial cells or vascular smooth muscle cells in vivo following myocardial damage. OBJECTIVE: To observe distribution, growth and differentiation of bone marrow mesenchymal stem cells (BMSCs) transplanted by different methods in vitals. METHODS: A total of 39 Japanese rabbits aged 3 months were selected to establish animal models of acute myocardial infarction, and then randomly divided into four groups. Three experimental groups were injected with BrdU-labeled autologous BMSCs through direct injection in myocardium, coronary artery injection and intravenous injection. The control group contained three subgroups, which were separately treated with an equal volume of saline through direct injection of coronary artery, vein and myocardium. At 4 weeks, animals were sacrificed to obtain heart, lung, liver and kidney to make tissue sections. BrdU-positive cells and BrdU- and α-striated muscle actin-positive cells were observed. RESULTS AND CONCLUSION: ①BMSCs labeled with BrdU and BrdU + α-striated muscle actin were found in and surrounding the infarct in each experimental group. Some of them had been connected with host myocardium and each other, with obvious transverse striation and sarcomere. ②The density of blood capillary increased in each experimental group compared with the control group (P < 0.05). The effectiveness sequence was coronary artery injection group, myocardium injection group and vein injection group from good to poor. ③The cells labeled with BrdU were found in the liver, lung and kidney of two groups which were transplanted through coronary artery and vein. Moreover, numbers of BrdU-positive cells were more in the lung than in the liver and kidney of the vein injection group. But, they did not differentiate and there were no tumor cells. Results have suggested that all three transplanting methods can induce regeneration of myocardial cells and blood capillaries in and surrounding the infarct region. Except myocardium injection group, BrdU-positive cells can be determined in non-target organs, but still mainly expressed in target organs. Related Articles | Metrics

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Effects of mobilization and collection of hemopoietic stem cells on normal donors Chen Xiao-xia, Wang Zhi-ming, Luo Xian-sheng, Xu Dan-dan, Li Xing, Lei Mei-qing 2010, 14 (23):  4281-4284.  doi: 10.3969/j.issn.1673-8225.2010.23.023 Abstract ( 467 )   PDF (380KB) ( 625 )   Save BACKGROUND: Mobilization of granulocyte colony-stimulating factor (G-CSF) is a main method used in healthy voluntary donor. Previous studies have demonstrated that some severe adverse effects occurred in mobilization of G-CSF. This leads to anxiety of the safety of unrelated healthy donors. Will the mobilization of granulocyte colony-stimulating factor have any effect on healthy voluntary donor? It remains unknown whether it is safe? OBJECTIVE: To investigate the effect of G-CSF on healthy donors. METHODS: A total of 16 healthy donors were mobilized using G-CSF at a dose of 5-10 μg/kg per day at Haikou People’s Hospital from January 2003 to December 2008. The adverse events were recorded during the process of mobilization and collection. Percentage of CD3, CD19, CD3+4, CD3+8 cells in blood was detected prior to and following mobilization. All donors were followed-up. RESULTS AND CONCLUSION: No adverse effects were determined during G-CSF mobilization in 10 donors. Three cases affected low-grade fever, headache, muscle and skeleton pain or lumbago. Three cases suffered from fever. Severity was in grade I, which does not cause the termination of the mobilization. Leukocyte number was greater following G-CSF mobilization than prior to mobilization. Leukocyte count returned to pre-mobilization levels at 3 days following stopping mobilization. There was no significant difference in hemoglobin and platelet counts, the percentage of CD3, CD19, CD3+4, CD3+8 cells in blood between pre-mobilization and 72 and 96 hours post-mobilization. Results have indicated that the healthy donors can tolerate the mobilization at a dose of 5-10 μg/kg per day, and G-CSF dose not affect the lymphocyte subsets. Related Articles | Metrics

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Effects of active principle region of Yangxing Tongmai Formula on bone marrow mesenchymal stem cell mobilization and homing in rats with acute myocardial infarction Li Yong-hua, Yuan Zhao-kai, Zheng Jing-hui, Wang Li-ping, Jian Wei-xiong, Huang Xian-ping 2010, 14 (23):  4285-4289.  doi: 10.3969/j.issn.1673-8225.2010.23.024 Abstract ( 351 )   PDF (573KB) ( 502 )   Save BACKGROUND: During acute infarct myocardium repair process, it is important to seek mobilization agent that can elevate number of peripheral blood bone marrow mesenchymal stem cells (BMSCs) and induce BMSCs homing to damaged myocardium. OBJECTIVE: To explore the effect of active principle region of Yangxin Tongmai Formula (apr-YTF) on the BMSC mobilization and homing of infarct myocardium in rats. METHODS: The apr-YTF was primarily composed of ginsenoside, tanshinone ⅡA, total alkaloids, and panaxan, which was supplied by College of Pharmacy, Central South University in accordance with the technique of this group. Among 70 Sprague Dawley rats, 8 rats were considered as normal control group, and the remaining 62 rats was used to establish models of acute myocardial infarction. A total of 32 rat models were randomly assigned to apr-YTF group, Danshen injection group, recombinant human granulocyte colony stimulating factor (rhG-CSF) group and model control group. After 3-hour modeling, corresponding drugs were given in each group for 5 days. Peripheral blood CD34+ cell number was determined using flow cytometry. CD34+ cell number was detected at marginal zone surrounding the infarct myocardium utilizing immunohistochemical staining. RESULTS AND CONCLUSION: Number of peripheral blood CD34+ cells increased significantly in the rhG-CSF group and apr-YTF group compared with model control group (P < 0.05). CD34+ cell number at marginal zone surrounding the infarct myocardium increased significantly (P < 0.01). Compared with the rhG-CSF group, CD34+ cell number at marginal zone surrounding the infarct myocardium increased significantly in the apr-YTF group (P < 0.01). The apr-YTF could promote BMSCs mobilization into blood, and directly homing to infarct myocardium. Its effects were identical to rhG-CSF. The effects on homing of CD34+ cells were better than rhG-CSF. It was a good agent for BMSCs mobilization. Related Articles | Metrics

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Primary culture of human periodontal ligament cells using three methods Jiang Jun-qiang, Wang Zhong-chao, Li Chun-hui, Cai Wei 2010, 14 (23):  4290-4294.  doi: 10.3969/j.issn.1673-8225.2010.23.025 Abstract ( 528 )   PDF (347KB) ( 576 )   Save BACKGROUND: The culture of human periodontal ligament cell (hPDLC) is the basis of biological research of periodontal cells. The success rate of primary culture of periodontal membrane cells is low, which restricts the biological research of periodontal cells. How to seek a simple effective culture method or how to elevate success rate of primary culture is always one of hot focus studies of biology of periodontal cells. OBJECTIVE: To explore an effective culture technique of hPDLC by comparing the effects of tissue explant, enzymatic digestion, the combination of tissue explant and enzymatic digestion. METHODS: The tissues of periodontal ligament isolated from 58 normal human permanent teeth were collected and divided into three groups randomly, and then primarily cultured using tissue explant, enzymatic digestion, the combination of tissue explant and enzymatic digestion, followed by subculture. Cell growth and morphology were observed under an inverted microscope. Vimentin and keratin were identified by immunohistochemistry. The fourth passage of cells were collected to draw growth curve. RESULTS AND CONCLUSION: The successful rate of the combination of tissue explant and enzymatic digestion (35%) was significantly higher than others (11% and 21%). The morphological and biological characteristics of the hPDLC in present study were similar to those typical hPDLC, which grew well. The growth curve of the fourth passage of cells was similar to “S” shape, with significant detention period, logarithmic phase and plateau phase. Results have suggested that the combination of tissue explant and enzymatic digestion is relatively simple and has a high successful rate, so it can be regarded as an effective method for hPDLC culture. Related Articles | Metrics

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Stable transfection of green fluorescent protein in rat fetal liver stem cells Yang Huan, Ye Zhang-qun, Chen Zhi-qiang, Wang Bo-han, Yao Wei-min 2010, 14 (23):  4295-4298.  doi: 10.3969/j.issn.1673-8225.2010.23.026 Abstract ( 314 )   PDF (521KB) ( 330 )   Save BACKGROUND: Fetus liver-derived liver stem cells, as cell transplantation donor, have advantages in source. However, difficult culture, easy to differentiate following passage and immature tracing localization technique restrict liver stem cell transplantation. OBJECTIVE: To observe whether primary cultured liver stem cells after tranfection by electroporation with pEGFP-C1 plasmid can stably express green fluorescent protein (GFP). METHODS: Mechanical separation and enzymatic digestion method were used to isolate liver stem cells from fetal Sprague Dawley rat liver tissues following pregnancy for 13.5 days. Half-amount medium replacement was used for purify isolated fetal liver cells following cultured in specific medium. Immunofluorescence technique was used to identify adherent cells. The plasmid pEGFP-C1 was identified correctly by eletrophoresis and then transfected into fetal liver stem cells． RESULTS AND CONCLUSION: The isolated fetal liver stem cells adhered to the culture plastic and presented round or oval cells after 24-hour cultivation in vitro. Isolated cells were almost identical circular after 2-day cultivation. At 3 days, cells grew into a colony which was constructed by 3 or 5 cells, with cell diameter of 6-10 μm; at 5 days, cell colony became enlarged and was composed of 10 dense, round cells of various sizes with clear boundary. At 8 days, they grew like epithelium cell. At 12 days, cells became big, extended like fried egg, with irregular forms, particles in cytoplasm, grew slowly. Following passage, there were no significant changes in cell amplification speed. Cells still presented epithelium-like shape at the passage 3. The adhered cells at day 5 following primary incubation were positively for human stem cell factor receptor CD34 and cytokeratin 19 using immunofluorescence technique. Green fluorescence was observed in many stem cells which has been transfected by pEGFP-Cl．This study successfully established liver stem cells with GFP. Related Articles | Metrics

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Isolation and culture of ICR mouse embryonic fibroblasts and production of feeder layers Fang Chi-hua, Hu Hai-bei, Hao Jian-zhi, Chen Xia 2010, 14 (23):  4299-4302.  doi: 10.3969/j.issn.1673-8225.2010.23.027 Abstract ( 511 )   PDF (439KB) ( 860 )   Save BACKGROUND: Mouse embryonic fibroblast (MEF) as feeder layer can be effective to promote embryonic stem cells and to induce pluripotent stem cell proliferation and to maintain their undifferentiated and pluripotent characteristics, which are better than feeder-free or adding single cytokines in medium, furthermore, the feeder layer can provide embryonic development environment for the above two kinds of cells. OBJECTIVE: To establish a stable and effective system of ICR MEF feeder layers to support embryonic stem cells and induce pluripotent stem cells in vitro. METHODS: MEF of ICR mouse was isolated using the method of trypsin digestion to observe effects of different mass concentrations of trypsin and different digestion time on the amount and the proliferation activity of MEF. Various concentrations of mitomycin C were utilized to treat MEF for 1, 1.5, 2, 2.5, 3, 3.5 hours to prepare feeder layer cells. MTT assay was employed to determine proliferation activity of MEF, and to explore optimal condition for preparing feeder layer. RESULTS AND CONCLUSION: The optimal mass concentration of trypsin for isolating MEF of ICR mouse was 0.05%; digestion time was 12-15 minutes; the optimal action mass concentration and time was 10 mg/L for 2.5 hours. Fibroblast feeder layer maintained for 8-12 days. The effects of 0.05% trypsin digestion for 15-20 minutes were superior to high mass concentration.  10 mg/L mitomycin C treated MEF for 2.5 hours can effectively inhibit the proliferation of the prepared feeder cells. Feeder layer cells prepared under this condition can support embryonic stem cells and induce pluripotent stem cells growth Related Articles | Metrics

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Preparation of mouse feeder layer cells and culture of mouse embryonic stem cell SF1-G Liu Feng, Lei Min-xiang, Chen Hui-ling 2010, 14 (23):  4303-4308.  doi: 10.3969/j.issn.1673-8225.2010.23.028 Abstract ( 353 )   PDF (561KB) ( 864 )   Save BACKGROUND: Mouse embryonic stem cell line SF1-G is derived from morula of female C57BL/6 mice mating with male M. spretus. Usually SF1-G was cultured on the layer of STO feeders. STO cells were expensive, whereas mouse embryonic fibroblasts (MEFs) were easier to get, and had more advantage in supporting embryonic stem cells growth, such as the formation of embryonic stem cell clones and maintaining the normal karyoplast of embryonic stem cells. Thus, it was very useful to establish a culture system in which SF1-G cells could amplify in an undifferentiated status. OBJECTIVE: To explore and establish an effective method of isolating and culturing MEFs and prepare the feeder cells for mouse embryonic stem cell (SF1-G cells) proliferation.  METHODS: The mouse primary MEFs were isolated from ICR mouse fetus at 12.5 to 14.5 day gestational ages. The 3 to 5 passages MEFs were treated by mytomycin C to inhibit cell proliferation. The treated MEFs were used as feeder cells for culturing SF1-G cell. The karyotype of SF1-G cell was detected by chromosome G staining process. The expression of alkaline phosphatase (AKP) of SF1-G cells was tested. Oct4 and Nanog gene expressions were also tested by RT-PCR. RESULTS AND CONCLUSION: MEFs were successfully isolated and cultured from mouse fetus. Feeder cells prepared MEFs of 3 to 5 passages could be used to support SF1-G cell growth with clear boundaries of clone. The SF1-G cells had normal karyotype, showing the positive results of AKP and expressing specific transcription factors. This experiment established an effective method of isolating and culturing MEFs and prepared feeder cells for propagating and culturing mouse embryonic stem cells, and SF1-G cells can be cultured in undifferentiated state in a laboratory. Related Articles | Metrics

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Culture of different sources-derived endothelial progenitor cells and comparison of their biological properties Xu Yan, Meng Heng-xing, Yu Zhen, Li Chang-hong, Qiu Lu-gui 2010, 14 (23):  4309-4316.  doi: 10.3969/j.issn.1673-8225.2010.23.029 Abstract ( 318 )   PDF (733KB) ( 687 )   Save BACKGROUND: Compensatory response in ischemic disease included arteriogenesis and angiogenesis. Endothelial progenitor cells (EPCs) play a key role in many physiological and pathological vascular remodeling. There are still controversies in the resource and biological characteristic of EPCs. OBJECTIVE: To isolate EPCs from peripheral blood, umbilical cord blood, bone marrow and umbilical cord, and to compare biological properties of the EPCs. METHODS: Mononuclear cells (MNCs) of peripheral blood, umbilical cord blood, and bone marrow were isolated by density-gradient centrifugation for adherent culture. Umbilical cord-derived EPCs were made into single cell suspension by adherent culture of explants or collagenase digestion of umbilical vein for adherent culture. The obtained adherent cells were detected for cell morphology, proliferation activity, cell cycle and immunophenotype. RESULTS AND CONCLUSION: ①Peripheral blood and umbilical cord blood-derived adherent cells grew as clones with spindle shape, and some round cells attached to them. With prolonged culture time, the round cells ablated gradually, and spindle cells had no appear proliferation. After digestion, these cells could not be subcultured. Flow cytometry showed that these cells highly expressed CD45 and partially expressed KDR. But they did not express CD31. ②Bone marrow-derived cells and umbilical cord-derived adherent cells were spindle and polygonal. Cells proliferated fast, and there was little change in cell morphology and proliferation activity after passage. Most cells were in G0/G1. Flow cytometry showed that these cells expressed neither CD14, CD45, nor CD106, HLA-DR, but highly expressed CD44, CD90, CD62E, CD73, CD95 and CD105. Some expressed endothelial marker: KDR, vWF. Umbilical cord-derived EPCs harvested by enzyme digestion were positive for CD31. Confocal microscope analysis revealed that all these cells could take up ac-LDL and bind UEA-1. Results have suggested that EPCs with different differentiation degrees and proliferation activities could be isolated from peripheral blood, umbilical cord blood, bone marrow and umbilical cord. Related Articles | Metrics

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Effects of physical therapy factors on mechanism of stem cell regulation Liu Ran, Fan Hong-xue, Liu Shi-wen 2010, 14 (23):  4319-4324.  doi: 10.3969/j.issn.1673-8225.2010.23.031 Abstract ( 397 )   PDF (578KB) ( 598 )   Save BACKGROUND: It is a hot focus on research for mechanism of stem cells modulated by chemical factors at present. But, many researches have shown that physical factors stimulation might also interfere in stem cell modulation to make target cell differentiation and generate function.   OBJECTIVE: To explore the application value of some physical factors on stem cell oriented differentiation.  METHODS: We reviewed about 102 research papers on common physical therapy factors on stem cell multiplication and differentiation in ten years. It was found that many researches much more concentrated on pulsed electromagnetic fields, transcranial magnetic stimulation, low frequency impulse electrotherapy, ultrasonic therapy, high pressure oxygen and various mechanical stress, which presumed that these physical factors might facilitate stem cell multiplication and differentiation, while few research on treadmill training and direct current stimulation showed that there might be no exact concern or report because of restriction on research thought or means. RESULTS OR CONCLUSION: The mechanism of physical factors interfering stem cells is complex. Physical factors might participate in cell growth microenvironment composed by biochemical environments including cell factors and growth factors and might have a supplementary or coordinated role in providing biochemical information such as cell factors. Related Articles | Metrics

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Research progress in islet stem cells: How far from the induction and production of functional beta cells？ Du Zhi-jian, Zhao Chun-hua 2010, 14 (23):  4325-4328.  doi: 10.3969/j.issn.1673-8225.2010.23.032 Abstract ( 373 )   PDF (463KB) ( 331 )   Save BACKGROUND: Insulin replacement therapy is the widely used method for diabetes treatment, whereas there are many defects for this treatment. Islets transplantation has been widely accepted as an effective way for diabetes treatment. But, transplantation rejection and shortage of cadaver pancreases limit its use in the clinic. Islet stem cells can solve this problem effectively. OBJECTIVE: To summarize source, induction, and differentiation efficiency of pancreatic β cells, present existing problems and application perspective. METHODS: We searched Pubmed database for relevant literatures concerning diabetes mellitus stem cells published from January 1990 to December 2009 using key words of “diabetes, stem cell, treatment, islets”. Following reading titles and abstracts, articles were primarily screened. Duplicated and Meta analysis studies were excluded, and 23 literatures were included for summary. RESULTS AND CONCLUSION: Pancreatic β cells derived from stem cells play a role in regulating blood glucose. At present, stem cells that can differentiate into pancreatic β cells primarily included embryonic stem cells, pancreatic stem cells, and bone marrow mesenchymal stem cells. In studies of differentiation of various stem cells into pancreatic β cells, there are two differentiation methods (in vivo and in vitro). In vitro induction mainly used step-by-stop induction, and some tests utilized gene technique. At present, studies addressing islet stem cells remain in a primary stage. How to induce and produce a large number of functional β cells is still a big challenge. Related Articles | Metrics

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Bone marrow mesenchymal stem cells transplantation for kidney diseases   Zhao Hong-mei, Hu Xiang, Li Fang, Jia Dan-bing 2010, 14 (23):  4329-4342.  doi: 10.3969/j.issn.1673-8225.2010.23.033 Abstract ( 395 )   Save BACKGROUND: Previous research has proved that bone marrow mesenchymal stem cells (MSCs) transplantation can repair injured kidney and the transplanted MSCs can differentiate into renal tubular epithelial cells, and thereby recovere the structure and function of kidney. OBJECTIVE: To analyze the application and the potential mechanisms of MSCs for treating kidney disease. METHODS: A computer-based search of Chinese and English Scientific and Technological Periodical Databases was performed to identify the articles about the correlation between MSCs and kidney disease, with the key words of “bone marrow mesenchymal stem cells, kidney disease”, and the language was limited to Chinese and English. After the first round of selection, the quotations of each literature were also reviewed, and those related to MSC transplantation and kidney disease and published in authoritative journals within 5 years were included. But repetitive articles and Meta analysis were excluded. RESULTS AND CONCLUSION: Transplantation of MSCs could promote the recovery of renal function by differentiating into glomerulus and tubular cells and it could also promote renal secretion. However, MSC transplantation for treating kidney disease still stayed at clinical test level, and animal models were the major research subjects. Moreover, clinical application for treating IgA kidney disease and secondary kidney disease, such as lupus nephritis, renal amyloidosis, and metastatic renal cell carcinoma, needs to be further studied. Related Articles | Metrics

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New direction of acute tubular necrosis treatment: Mechanism of promoting kidney repair by exogenous mesenchymal stem cells Liu Lin, Cai Guang-yan, Zhang Li 2010, 14 (23):  4333-4336.  doi: 10.3969/j.issn.1673-8225.2010.23.034 Abstract ( 426 )   PDF (456KB) ( 445 )   Save BACKGROUND: Mesenchymal stem cells have become a spotlight in the field of tissue repair and regeneration, due to its extensive capacity of differentiation and secretion as well as simple collection, culture advantages and stable characteristics. In the field of nephrology, the ability of exogenous mesenchymal stem cells to promote the repair process following acute tubular necrosis has been proved in multiple animal researches. Meanwhile, researchers are making efforts to explore the exact mechanism. OBJECTIVE: To review current animal researches about exogenous mesenchymal stem cells on promoting the repair process of acute tubular necrosis, and to analyze the results to further elucidate the mechanism of this promoting effect. METHODS: We checked PubMed database about the related articles published from January 2004 to July 2009 on computer with key words of “mesenchymal stem cell, acute tubular necrosis, acute kidney injury, immune, cytokine, renal progenitor cells, tubular epithelial cells”. The language was limited to English. Simultaneously, we also checked the book Chinese Journal of Nephrology by hand. A total of 309 articles have been attained, with 306 from computer and 3 from the book, and 37 articles were consistent with inclusion criteria. RESULTS AND CONCLUSION: In the field of nephrology, a great number of animal researches have proved that exogenous mesenchymal stem cells could promote the repair process after acute tubular necrosis, which provides a prospective effect for the treatment of acute tubular necrosis. Up to now, the possible mechanisms of this process include: ①mesenchymal stem cells differentiate into tubular epithelial cells directly to renew the damaged structure. ②Mesenchymal stem cells modulate immune system and activate intrinsic kidney cells via endocrine or paracrine to help repair the injured tissue. Related Articles | Metrics

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Stem cell therapy for type 1 diabetes: Can it be an effective clinical scheme? Jing Hua, Zhang Jin-yuan, Lü Xu-jing 2010, 14 (23):  4337-4341.  doi: 10.3969/j.issn.1673-8225.2010.23.035 Abstract ( 721 )   PDF (506KB) ( 641 )   Save BACKGROUND: How to cure type 1 continuous development diabetes completely has received wide attention in the medical field all over the world. Recently, with the understanding of stem cells and the continuous development of the stem cell therapy for type 1 diabetes has become a research hotspot. OBJECTIVE: To review the research progress addressing stem cells for type 1 diabetes. METHODS: A computer based search was conducted in PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) for relevant articles published from January 1997 to June 2009, with key words “type 1 diabetes, embryonic stem cells, pancreatic stem cells, mesenchymal stem cells, hematopoietic stem cells” in English. Simultaneously, an additional research was performed in Chinese Journal Full-text database (http://www.wanfangdata.com.cn) for articles published from January 1997 to June 2009 with the key words of “type 1 diabetes, embryonic stem cells, pancreatic stem cells, mesenchymal stem cells hematopoietic stem cells” in Chinese. A total of 134 documents were retrieved. RESULTS AND CONCLUSION: Type 1 diabetes is T cell-mediated destruction of pancreatic β cells in autoimmune diseases, which needs exogenous insulin to control blood sugar. There is no cure for it. Stem cells are a class of pluripotent cells and can be induced to differentiate into insulin-producing cells, and stem cells have become the search for alternatives to β-cells induced by the new resources. Currently, stem cell research can be divided into embryonic stem cells and adult stem cells. Embryonic stem cells are mostly studied in animals, and proliferation and differentiation under in vivo environment are not easily controlled. Embryonic stem cells are derived from embryos, so ethics is not widely recognized, which limits clinical research. Adult stem cell research mainly addresses pancreatic stem cells, mesenchymal stem cells and hematopoietic stem cells. Pancreatic stem cell has unclear surface markers, and is characterized by difficult harvest, not easy to capture and difficult purification in the clinic. Mesenchymal stem cells are rich to be harvested, but its cross-mesoderm differentiation is still controversial, and the function maintenance time of the differentiated cells in vivo is uncertain. Hematopoietic stem cell, the most mature adult stem cells, is of the most extensive application; it is not only from rich source, easy to collect, but also inhibits rejection, has wide application and will be of a hot focus in type 1 diabetes research. Related Articles | Metrics

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Research and application of stem cell transplantation for treatment of heart disease Li Mu, Song Jian-fei, Liang Yue-pei, Li An-gui, Wang Wei 2010, 14 (23):  4342-4346.  doi: 10.3969/j.issn.1673-8225.2010.23.036 Abstract ( 295 )   PDF (501KB) ( 320 )   Save BACKGROUND: With the development of end-stage heart disease, heart disease can not be cured fundamentally due to lots of limitations and existing treatments. Stem cells are mature cells of various origin cells, with self-renewal and differentiation potential. Stem cells have a strong regenerative capacity to replace, repair or strengthen damaged tissues or organs of biological functions. In theory, it is possible to cure cardiovascular disease eventually. However, numerous studies have suggested that stem cell transplantation treatment is safe and effective. OBJECTIVE: To summarize the clinical research progress in stem cell transplantation in treatment of heart disease. METHODS: The first author retrieved PubMed Database (http://www.ncbi.nlm.nih.gov/PubMed) for related articles published from January 1999 to October 2006, with the key words of “stem cells, transplant, heart disease” in English. Simultaneously, a computer-based search was performed in China Journal Full-text Database (http://www.wanfangdata.com.cn) for relevant articles published from January 1999 to October 2006 with the key words of “stem cells, transplant, heart disease” in Chinese. A total of 60 literatures were retrieved. RESULTS AND CONCLUSION: Various types of heart disease developed to the final stage, the healthy myocardial cells cannot satisfy the needs of the body due to irreversible damage to the structure and function of myocardial cells. Moreover, varied palliative conservative treatments can not achieve satisfactory degree due to inability to solve the fundamental problem. However, the heart transplantation has been greatly limited, due to difficulties in obtaining donor, immunological rejection, high cost and high risk. These provide challenges for clinical treatment. Looking for a more effective treatment and opening up new treatment approaches have become inevitable. With the development of tissue and cell engineering research, the function recovery and reconstruction of stem cell transplantation therapies have become the hot research prospect. Related Articles | Metrics

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Adult neural cell regeneration after traumatic brain injury Zhao Wang-miao, Huan Lin-chun, Yang Xin-yu 2010, 14 (23):  4347-4350.  doi: 10.3969/j.issn.1673-8225.2010.23.037 Abstract ( 442 )   PDF (384KB) ( 366 )   Save BACKGROUND: Traumatic brain injury can lead to serious neurological dysfunction. Basal research reveals that persistent endogenous neurogenesis exists in adult mammal brain, which may benefit for recovery of brain trauma. OBJECTIVE: This article reviews some progresses of neurogenesis in the adult hippocampus, subventricular zone and cerebral cortex after traumatic brain injury to find effective measures to improve neurogenesis and restoration of neurological function. METHODS: We searched the related articles with respect to adult neurogenesis after traumatic brain injury in Pubmed database, published from January 1998 to March 2010. The key words were “traumatic brain injury, neurogenesis, hippocampus, subventricular zone, cerebral cortex” in title/abstract field. The content of articles was related to trauma and adult neurogenesis, and the study area involves neurogenic region (hippocampus and subventricular zone) and nonneurogenic region cerebral cortex. The articles which are newly issued or published in authoritative magazines were selected. In total 213 articles were included in initial search and 33 articles were selected to review according to inclusion criteria. RESULTS AND CONCLUSION: Neurogenesis existed in the adult nervous system. Moderate brain trauma could stimulate neurogenesis of hippocampus and subventricular zone, and neurogenesis contributed to restoration of hippocampal function. Some exogenous factors which can promote neurogenesis can also improve neurological function. Resting neural progenitor cells exist in the cerebral cortex. The latent cells will re-entry the cell cycle and induce neurogenesis under certain conditions and they may benefit for the recovery of brain trauma. Related Articles | Metrics

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Role of Notch signal transduction pathway in never repair and regeneration Wang Kai, Zhao Bin, Wang Shuan-ke, Wang Na 2010, 14 (23):  4351-4354.  doi: 10.3969/j.issn.1673-8225.2010.23.038 Abstract ( 286 )   PDF (448KB) ( 631 )   Save BACKGROUND: Neural stem cells differentiate into functional neurocytes though equity cleavage, thereby recovery function of damaged cells. Notch signal transduction determines neurons differentiating into neurons and gliocytes. OBJECTIVE: To summarize Notch structure, adjusting mechanism, influence factors, and effect on neurocytes. METHODS: Articles addressing signal transduction, differentiation and development of neural stem cells, and spinal cord injury were retrieved from CNKI and PubMed database with the key words of “Notch, neurocyte, neural repair, neural regeneration” or “signal transduction, NSC, SCI, Notch”. A total of 430 articles were retrieved, and 31 ones were included in the final analysis. RESULTS AND CONCLUSION: Effect of Notch on nerve repair and regeneration were mainly discussed. The Notch could selectively activate multi-functional neural stem cells in vitro and embryonic prosencephalic radiated neuroglia in vivo; moreover, activation is necessary for star-like neuroglia during the development, because it played an important role in adjusting development, growth, and apoptosis of other functional cells. Related Articles | Metrics

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Autologous bone marrow cell transplantation via spleen artery in 5 patients with type 2 diabetes mellitus Wang Pan, Wu Zheng-rong, Tian Jing-lun, Qi Long 2010, 14 (23):  4355-4358.  doi: 10.3969/j.issn.1673-8225.2010.23.039 Abstract ( 543 )   PDF (298KB) ( 617 )   Save BACKGROUND: Recently, more and more people try to use stem cells to treat diabetes mellitus. Application of autologous bone marrow stem cells is a simplest and safest method, but no reports have addressed autologous bone marrow stem cells for type 2 diabetes mellitus. There are several reports concerning the outcomes of diabetes mellitus treatment. Thus, there is no final conclusion at present. OBJECTIVE: To observe effectiveness of autologous bone marrow stem cells for type 2 diabetes mellitus via splenic artery transplantation, and to explore its feasibility, safety and validity. METHODS: A total of 5 patients with acute myocardial infarction and type 2 diabetes mellitus had been enrolled at the ICU Department, Wenjiang People’s Hospital from March to July 2009. They received insulin combined with oral administration of sugar-reducing drugs, but blood sugar was not fully controlled, and then subjected to autologous bone marrow cell transplantation. ①Patients received multipoint puncturation in the bilateral posterior superior iliac spine in a prone position following local anesthesia. A total of 100-150 mL bone marrow blood was collected. This test was accomplished within 60 minutes. Bone marrow cells were harvested from bone marrow blood, washed, and then diluted using saline into mononuclear cell suspension about 20-30 mL. ②Through femoral artery intubation, 4.0 cm judkins 5 or 6F right coronary arteriography catheter was inserted into the celiac trunk. The catheter direction was regulated, and the catheter entered splenic artery. Following injection of 5-10 mL ultravist, the catheter position was determined, and bone marrow cell suspension was inserted. ③Blood glucose levels and insulin dosage were followed up at 3 and 9 months following transplantation. RESULTS AND CONCLUSION: A total of 5 patients with coronary artery disease and type 2 diabetes mellitus received successfull transplantation and were followed up. ①During autologous bone marrow cell transplantation, no adverse reaction was detected; two cases developed abdomen fever; one case suffered from light nausea. ②Compared with pretransplantation, fasting blood glucose was reduced by 23% and 27% at 3 and 9 months after the transplantation. ③Compared with pretransplantation, daily insulin requirement was significantly reduced by 22% and 42% at 3 and 9 months after the transplantation. These results have demonstrated that selective spleen arterial transplantation of autologous bone marrow cells is safe and can decrease blood glucose level and daily insulin dose in patients with type 2 diabetes mellitus. Related Articles | Metrics

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Wnt3a induces rat bone marrow mesenchymal stem cells differentiation into neuron-like cells Wang Xiao-mei, Mu Chang-zheng, Ma Yun-sheng 2010, 14 (23):  4363-4366.  doi: 10.3969/j.issn.1673-8225.2010.23.041 Abstract ( 383 )   PDF (417KB) ( 443 )   Save BACKGROUND: Wnt signaling pathway is a key regulator of cellular proliferation and differentiation, but its correlation with neural differentiation of bone marrow mesenchymal stem cells (BMSCs) is not very clear. OBJECTIVE: To find out the molecules of the Wnt family which are involved in differentiation of rat BMSCs into neuron-like cells. METHODS: The rat BMSCs were separated and cultured in vitro. The morphology of the BMSCs was observed. Flow cytometry analysis was performed to detect cell phenotype CD44, CD9, CD34 and CD45. Wnt3a and Wnt5a were respectively combined with basic fibroblast growth factor to induce BMSCs differentiation into neuron-like cells, and then were identified by using immunocytochemistry and RT-PCR. RESULTS AND CONCLUSION: The BMSCs were long-spindle. CD9 and CD44 were highly expressed, while CD34 and CD45 were lowly expressed. Nestin and neuron specific enolase were positive but glial fibrillary acidic protein were not obviously expressed when they were cultured with Wnt3a. In Wnt5a group, Nestin expression was weakly positive, while neuron specific enolase and glial fibrillary acidic protein were negative. RT-PCR result revealed Nestin expressed both before and after induction in the Wnt3a induced group, neuron-specific enolase exhibited apparent amplified bands 5 days after the induction, and more apparent at 10 days. A weak amplification band of glial fibrillary acidic protein could be seen at 10 days after the induction. In Wnt5a and control groups, BMSCs induced by 10 days weakly expressed Nestin, while neuron-specific enolase and glial fibrillary acidic protein were almost not expressed. It is indicated that Wnt3a molecule can promote the differentiation of BMSCs cultured in vitro to neuron-like cells. Related Articles | Metrics

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Stem cells in optic nerve protection and restoration following injury Zhang Min, Huang Bing 2010, 14 (23):  4367-4370.  doi: 10.3969/j.issn.1673-8225.2010.23.042 Abstract ( 379 )   PDF (236KB) ( 540 )   Save BACKGROUND: As a specially differentiated central nerve, optic nerve is once injured, retinal neurons are hard to restore and regenerate. As a result, visual function is lost permanently. Currently, an effective clinical treatment is not available. Due to the multi-directional differentiated potential of stem cells, their role in optic nerve protection and restoration following injury is becoming a hot research spot. OBJECTIVE: To review the classification and transplantation of stem cells in optic nerve protection study, as well as roles of stem cells transplantation in optic nerve protection and repair following injury. METHODS: The database of ISI Web of Knowledge platform was searched on computer from February 1994 to April 2009, with the key words “stem cells, optic nerve” in English. In the same way, the database of CNKI from February 1994 to April 2009 was also screened, with the key words “stem cells, optic nerve” in Chinese. RESULTS AND CONCLUSION: A total of 98 literatures related to stem cells in optic nerve protection and restoration after injury are collected, in which 23 papers are Chinese and 75 papers are English. The studies published earlier, repetitively and similarly are removed, and 38 papers which comply with the standards are involved into this review. Stem cells own the potential of self-renew physiologically, as well as potential of repairing the injuries. At present, most of the researches on stem cells in optic nerve protection and restoration after injury are represented in morphosis. How to implant stem cells safely and effectively and how to recover stable visual function still need our efforts. Stem cells derived from skin, peripheral blood, fat and other tissues are convenient to draw materials and are commendable subjects, we should study them deeply to make preferable use. Related Articles | Metrics

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Autologous bone marrow mesenchymal stem cells transplantation for the treatment of severe diabetic foot in one case Chen Ling-zhen, Chen Jia-yu, Wu Jin-ming, Yu Wei, Zhan Yu 2010, 14 (23):  4539-4362.  doi: 10.3969/j.issn.1673-8225.2010.23.040 Abstract ( 315 )   PDF (505KB) ( 509 )   Save BACKGROUND: Previous studies have shown that bone marrow mesenchymal stem cells (BMSCs) can differentiate into vascular endothelial cells, and then further differentiate into new blood capillary, resulting in forming new collateral branch and in improving blood supply in local site, which can provide new ideas for treatment of ischemic lower extremity disorders. OBJECTIVE: To observe clinical effects of autologous BMSCs transplantation on the treatment of severe diabetic foot. METHODS: One patient with type 2 diabetes and severe diabetic foot ulcers was included in this research. BMSCs were isolated and expanded from 50 mL bone marrow aspirates through lower-extremity artery CT vascular reconstruction, which suggested lower-extremity angiosclerosis, stenosis and occlusion. Mononuclear cells were harvested by density gradient centrifugation, and BMSCs were isolated and purified using adherent culture method, and then infused into patients via intravenous infusion following microbiological detection. At 3 and 6 months, 7 and 30 days following transplantation, clinical symptoms such as pain, cold sensation, intermittent claudication, ankle-brachial index (ABI), ulcer area and gangrene were observed. Improvement and blood flow in the lower extremity and artery lateral branch reconstruction were measured by color Doppler flow imaging and CT scan. RESULTS AND CONCLUSION: At 3 months following transplantation, pain and cold sensation significantly relieved and numbness was improved. At 6 months after transplantation, pain relief was achieved, ABI significantly increased, foot ulcers healed completely, lower extremity was maintained, and foot function in walking was well preserved. New vessels formation was visible and blood flow was significantly improved by color Doppler flow imaging and CT scan. No heart/live/lung/kidney function injury was observed during the transplantation. The patient was followed up for 14 months, no transplantation-related complications were observed. Autologous BMSCs transplantation provides a new way for the therapy of diabetic foot. Related Articles | Metrics