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    03 December 2011, Volume 15 Issue 49 Previous Issue    Next Issue
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    Biological characteristics of bone marrow mesenchymal stem cells derived from coagulation factor Ⅸ knockout mice
    Lü Min-min, Lin Hong-gang, Zhou Guang-qian
    2011, 15 (49):  9121-9127.  doi: 10.3969/j.issn.1673-8225.2011.49.001
    Abstract ( 335 )   PDF (2156KB) ( 323 )   Save

    BACKGROUND: Coagulation factor Ⅸ knockout may affect the biological characteristics of bone marrow mesenchymal stem cells (BMSCs) of the bone marrow microenvironment by affecting the hematopoietic system stability.
    OBJECTIVE: To characterize the biological properties of BMSCs derived from coagulation factor Ⅸ knockout mice.
    METHODS: BMSCs were isolated from coagulation factor Ⅸ knockout mice C57BL/6J-F9 tm1. Dws by using a modified adherent culture method. The cells were detected for their expression of general MSCs surface markers by using flow cytometry. The functionality was assessed for their multipotency to differentiate into osteoblasts, adipocytes and chondrocytes upon certain induction conditions.
    RESULTS AND DISCUSSION: The BMSCs showed spindle-shape morphology and high capacity for proliferation and self-renewal. The cells were positive for CD44, CD56, CD73, CD90, CD105, CD106, CD166 and CD271, but negative for CD34 and CD11b. They demonstrated the ability to differentiate into osteoblasts, adipocytes and chondrocytes. These results illustrate the BMSCs derived from coagulation factor Ⅸ knockout mice C57BL / 6J-F9 tm1. Dws have perfect stem cell biological characteristics. The results initially suggest that deficiency of coagulation factor Ⅸ has no effect on the biological characteristics of BMSCs.

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    Chondrometaplasia of bone marrow mesenchymal stem cells of different resources in vivo
    Pan Xin-yu, Cui Ying, Ma Lin-xiang, Wang Yue-tian, Yuan Yang, Wang Xue-feng
    2011, 15 (49):  9128-9132.  doi: 10.3969/j.issn.1673-8225.2011.49.002
    Abstract ( 323 )   PDF (1704KB) ( 487 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can repair cartilage defects after induction in vitro. However, most of the seed cells come from autos or allograft currently.
    OBJECTIVE: To observe the effects of the chondrometaplasia of homologous and heterogeneous BMSCs on repairing the laryngeal cartilage defects.
    METHODS: The third generation of human embryonic BMSCs and rabbit BMSCs were involved. The two kinds of cells grew on poly lactic acid-glycolic acid scaffold, and were inducted to cartilage cells after adding transforming growth factor β1 and cartilage morphogenetic proteins into the scaffold. The two kinds of cell systems were implanted in New Zealand rabbits. The defects were removed at 4 and 8 weeks after implantation for general and histological observation.
    RESULTS AND CONCLUSION: The human embryonic BMSCs and rabbit BMSCs were filled with regenerated tissues at 4 and 8 weeks after implantation. Histological observation showed that most cells in the tissues were cartilage cells which could secrete glycosaminoglycan and collagen Ⅱ. The two types of cell scaffold could generate cartilage cells in about the same amount, and there was no immunological rejection between them. Results revealed that the chondrometaplasia of the heterogeneous BMSCs induced by transforming growth factor β1 and cartilage morphogenetic proteins is similar to the homologous BMSCs for repair of the laryngeal cartilage defects.

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    Dose-dependent effect of erythropoietin on the proliferation of rabbit bone marrow mesenchymal stem cells
    Li Zheng-zhang, Shen Zhe, Cheng Ying-zhang, Xue Hao-ping, Ji Qun, Cheng Shi, Xue Min-zhi
    2011, 15 (49):  9133-9136.  doi: 10.3969/j.issn.1673-8225.2011.49.003
    Abstract ( 385 )   PDF (1233KB) ( 338 )   Save

    BACKGROUND: Some studies have certificated that erythropoietin (EPO) can promote bone marrow mesenchymal stem cells (BMSCs) proliferation, but it is not yet clear about the dose-dependent relationship.
    OBJECTIVE: To explore the different-dose effects of EPO on rabbit BMSCs proliferation activity and cell cycle.
    METHODS: MTT colorimetric test and flow cytometry were used to detect the effect of EPO (0, 2, 4, 8, 16 U/mL) on BMSCs cell cycle; meantime, the culture medium was collected for matrix metalloproteinase-2 (MMP-2) content detection.
    RESULTS AND CONCLUSION: 2-16 U/mL EPO could promote BMSCs proliferation gradually in a dose-dependent manner. BMSCs percentage in S phase and proliferative index gradually increased with the dose increase of EPO, and the proliferation index of BMSCs with EPO was significantly higher than that of BMSCs without EPO (P < 0.05); however, the level of MMP-2 was decreased gradually, especially in 16 U/mL EPO group. These indicate that EPO can increase the proliferative ability of BMSCs in a dose-dependent manner.

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    Effects of two separation methods on biological characteristics of sheep bone marrow mesenchymal stem cells
    Tang Xiao-xue, He Hui-yu, Xu Hui-fen, Hu Yang, Xu Guo-qiang
    2011, 15 (49):  9137-9140.  doi: 10.3969/j.issn.1673-8225.2011.49.004
    Abstract ( 249 )   PDF (1337KB) ( 361 )   Save

    BACKGROUND: In recent years, there are numerous studies on bone marrow mesenchymal stem cells (BMSCs) from small animals such as rats and rabbits, but reports addressing BMSCs from large animals, such as sheep, are rare.
    OBJECTIVE: To observe the effect of two isolated methods on the biological activity of sheep BMSCs.
    METHODS: Fresh bone marrow of 10 mL was extracted from the posterior superior iliac spine of a healthy Altay sheep aged two months by puncture following anesthesia, and BMSCs were obtained by the whole bone marrow culture method and density gradient centrifugation method. The shape and adherent growth of primary BMSCs was observed under inverted fluorescence microscope. Primary culture time in two groups was recorded, and the CD44 antigen expression was identified by flow cytometry. Cell growth curve was measured by MTT assay. The potential of osteogenic differentiation was examined by Von Kossa’s staining.
    RESULTS AND CONCLUSION: Using both of these two separation methods, more uniform spindle, triangle and polygon BMSCs could be harvested, the nuclei were big and cytoplasm was rich, which were positive for CD44 antigen. The whole marrow culture was better than the density gradient centrifugation in cells quantity, slightly early in culture time. Von Kossa staining was positive in the two groups after BMSCs were cultured for 14-21 days. The adherent growth of passaged BMSCs obtained by using the whole bone marrow culture method was faster than that using the density gradient centrifugation culture method, and the cell vitality was also better in the former. It is indicated that the whole marrow culture is an effective method of extraction.

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    Bone marrow mesenchymal stem cells modified by glial cell line-derived neural factor gene co-cultrued with neurons subjected to anoxia-reoxygenation
    Xiong Huai-lin, Deng Li, Gao Xiao-qing, Wang Te-wei, Guo Kan, Yang Chao-xian
    2011, 15 (49):  9141-9145.  doi: 10.3969/j.issn.1673-8225.2011.49.005
    Abstract ( 272 )   PDF (1498KB) ( 388 )   Save

    BACKGROUND: Preliminary studies in workgroup indicated that glial-derived neurotrophic factor (GDNF) gene modified by bone marrow mesenchymal stem cells (BMSCs) (BMSCs/GDNF) can stably express the mRNA of GDNF, but whether GDNF protein in BMSCs/GDNF affects neurons subjected to anoxia-reoxygenation positively is unclear.
    OBJECTIVE: To observe the protective effects of BMSCs/GDNF on neurons subjected to anoxia-reoxygenation.
    METHODS: BMSCs/GDNF were co-cultured with anoxia-reoxygenation nerve cells. Hoechst33258 staining was used to detect apoptosis of neurons. Immunofluorescent-chemistry method was used to detect growth-associated protein-43 (GAP-43) expression in co-cultured cells.
    RESULTS AND CONCLUSION: The rate of neurons apoptosis in the BMSCs/GDNF group was lower than that of the BMSCs group and anoxia group (P < 0.05). Absorbance of growth-associated protein-43 (GAP-43) expression in the BMSCs/GDNF group was higher than that in the BMSCs group (P < 0.05). It is indicated that BMSCs/GDNF have neuroprotective effects due to inhibiting apoptosis of neurons subjected to anoxia-reoxygenation and increasing GAP-43 expression in co-cultured cells.

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    Expression of albumin and cytokeratin 18 in human umbilical cord blood mesenchymal stem cells induced by serum of patients with server liver diastase
    Zhao Wen-jing, Sun Guo-jun, Chen Xi, Wang Jia-nan, Liu Shu-rong, Ju Hong-mei
    2011, 15 (49):  9146-9150.  doi: 10.3969/j.issn.1673-8225.2011.49.006
    Abstract ( 378 )   PDF (1587KB) ( 339 )   Save

    BACKGROUND: Studies have found that the cord blood contains a large amount of mesenchymal stem cells (MSCs). These cells have quicker speed of getting into cell cycle and stronger autocrine ability of growth factors than those in the bone marrow or peripheral blood after activation.
    OBJECTIVE: To investigate the differentiation ability of human umbilical cord blood MSCs (HUMSCs) by serum of patients with severe liver disease.
    METHODS: HUMSCs were isolated and purified by density gradient centrifugation and adherent culture. HUMSCs were induced to differentiate with 20% serum of liver failure patients. On days 7, 14, 21 after induction, the expressions of albumin and cytokeratin 18 were identified by immunohistochemical method.
    RESULTS AND CONCLUSION: High purity HUMSCs were isolated from umbilical cord blood. HUMSCs strongly expressed CD44 and CD29, but did not express CD34. HUMSCs may differentiate into adipocytes in vitro. Morphological changes happened in HUMSCs cultured in L-DMEM medium containing 20% serum of patients with severe liver disease, the cells became larger and flat showing epithelioid cells type. Streptavidin-perosidase showed that only few cells were positive for albumin and cytokeratin 18 in the experimental group after 7 days induction. The positive rates of albumin and cytokeratin 18 were increased after 14 and 21 days induction, which were significantly different between the experimental group and control group (P < 0.05). It is indicated that high purity HUMSCs can be isolated by density gradient centrifugation and adherent culture, and serum of liver failure patients can induce the albumin and cytokeratin 18 expression in HUMSCs.

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    Optimizing transfection of human umbilical cord-derived mesenchymal stem cells using novel chemical transfection reagents
    Lu Jun, Zhao Hong-zhou, Chen Jin, Fu Yun-feng, Huang Liang-hu, Tan Jian-ming
    2011, 15 (49):  9151-9154.  doi: 10.3969/j.issn.1673-8225.2011.49.007
    Abstract ( 320 )   PDF (987KB) ( 500 )   Save

    BACKGROUND: Genetic modification technology of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) is numerous and varies in the complexity of their efficiency, reliability and safety.
    OBJECTIVE: To compare transfected efficiency of different novel chemical transfection reagents on hUCMSCs.
    METHODS: The hUCMSCs were transfected with Fugene HD, Lipofectamine LTX, and Attractene. Positive cells were detected under fluorescent microscope, and expression efficiencies were measured by using flow cytometry. Cell viability was measured by using trypan blue exclusion after transfection.
    RESULTS AND CONCLUSION: The highest transfected efficiency was observed with Lipofectamine LTX (32.5±2.12)% (P < 0.05), followed by Fugene HD (4.3±0.64)%, minimal transfection efficiency was observed with Attractene (1.74±0.08)%. Lipofectamine LTX demonstrated the lower cell viability (69.8±6.3)% compared with Fugene HD (92.4±4.2)% and Attractene (106.6±3.9)% (P > 0.05), but the difference was not significant. In summary, this study demonstrates that Lipofectamine LTX is a relative ideal chemical transfection reagent for hUCMSCs.

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    Safety of umbilical cord versus bone marrow-derived mesenchymal stem cells transplantation for the treatment of nervous system lesions
    Huang Huai, Chen Hui-qiang, Pang Yan, Gu Jing, Yu Rong-hao, Xiao Yang
    2011, 15 (49):  9155-9159.  doi: 10.3969/j.issn.1673-8225.2011.49.008
    Abstract ( 286 )   PDF (436KB) ( 305 )   Save

    BACKGROUND: The comparative study concerning the safety of umbilical cord and bone marrow-derived mesenchymal stem cells transplantation for the treatment of nervous system lesions is insufficient. 
    OBJECTIVE: To assess the safety of umbilical cord and bone marrow-derived mesenchymal stem cells transplantation for treatment of nervous system lesions.
    METHODS: A total of 214 cases with neuropathy were randomly divided into A, B groups. Patients in the A group received umbilical cord derived stem cell transplantation, and those in the B group received bone marrow-derived mesenchymal stem cells transplantation. Totally (5-12)×108 stem cells were transplanted into each patient.
    RESULTS AND CONCLUSION: The count of lymphocytes, alanine aminotransferase, aspartate aminotransferase, IgA, and IgM were increased compared with those before treatment in both groups (P < 0.01); However there were no significant differences between two groups (P > 0.05). Moreover, white blood cell count and red blood cell count in cerebrospinal fluid of all patients were significantly greater than the normal level. There were no significant differences between two groups (P > 0.05). No significant differences of the positive rate of Pandy test and the incidence rate of adverse effect were found in both groups (P > 0.05). The safety of umbilical cord and bone marrow-derived mesenchymal stem cell transplantation for treatment of nervous system lesions showed no marked differences.

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    Guinea pig adipose-derived mesenchymal stem cells: Ideal stem cells for experimental studies of otology
    Guo Wei, Ma Yan, Duan Li-fu, Bi Xiao-juan, Liu Li-zhong
    2011, 15 (49):  9160-9164.  doi: 10.3969/j.issn.1673-8225.2011.49.009
    Abstract ( 373 )   PDF (1580KB) ( 285 )   Save

    BACKGROUND: Up to date, most of the animal experiments about stem cells research on the otology use the stem cells of other animal species for the guinea pig model, however, the research of guinea pig-derived stem cells is rarely reported.
    OBJECTIVE: To explore the differentiated potential of the guinea pig adipose mesenchymal stem cells into osteoblasts, adipocyte, nerve cells, and the application possibility for experimental study of otology.
    METHODS: Guinea pigs adipose tissue was isolated and cultured to the 3rd generation. Absorbance values of cell proliferation from 1 to 10 days were detected and the immunophenotype of the cells were detected by flow cytometry. Differentiation of osteoblast, adipocyte and nerve cells were verified.
    RESULTS AND CONCLUSIONS: The growth curve of adipose mesenchymal cell proliferation is similar to S-shaped, the first 3 days was the incubation period after passage and the cell populations entered logarithmic growth phase from the 4th day, after the 8th day they entered into the platform period. The rejection rate of trypan blue staining was 94%-98%, the total cells was amplified by about 3 600 times from primary to passage 5, The expression rate of cells of the immune phenotype CD29 was 86.3%, CD44 was 55.5% and CD45 was 0.4%. After differentiation of osteoblast, specific staining demonstrated the formation of mineralized nodules. After differentiation of adipocyte, lipid droplets could be seen in cytoplasm by oil red O staining. After differentiation of nerve cells, cells morphology was similar to neural cells and the results of Nestin, GFAP, NF200 staining were positive. Guinea pig adipose mesenchymal stem cells was easy to isolate, culture and amplification in vitro. They have the “stem” characteristics of mesenchymal stem cells, and they were ideal stem cells for experimental studies of otology.

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    Effect of X box-binding protein 1 on the proliferation of endogenous neural stem cells after focal cerebral ischemia
    Xing Xue-song, Lü Wei-li
    2011, 15 (49):  9165-9168.  doi: 10.3969/j.issn.1673-8225.2011.49.010
    Abstract ( 224 )   PDF (1115KB) ( 296 )   Save

    BACKGROUND: Nuclear transcription factor X box-binding protein 1 (Xbp1) is expressed during the development of the central nervous system, and plays an important role in neurons cell proliferation.
    OBJECTIVE: To investigate the expression of nuclear transcription factor Xbp1 in neural stem cells of cerebral ischemia/reperfusion hippocampal tissue and the inhibition effects of basic fibroblast growth factor (bFGF). 
    METHODS: A novel model of cerebral ischemia/reperfusion was established with the method of middle cerebral artery occlusionin rats. The rats were divided into control group, ischemia/reperfusion group and bFGF group. The expression of BrdU and Xbp1 and the inhibition effects of bFGF in hippocampal neural stem cells were detected with immunohistochemical method.
    RESULTS AND CONCLUSION: After 3 days, BrdU positive cells in the hippocampus were obviously increased. To the 7th day, BrdU positive cells were more than those at any time. At 3 days, Xbp1 reaction products in ischemia/reperfusion were more than that in control group, and reached the peak on day 7, and then gradually decreased. bFGF can promotes the expression of Xbp1 in the hippocampus tissue and the proliferation of neural stem cells. These findings indicate that bFGF promotion of neural stem cells proliferation may be mediated by Xbp1 signaling pathway.

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    Development of current mediated by NMDA receptors during differentiation of neural stem cells into neuronal precursor cells
    Wang Xiang-zhen, Li Mei, Hu Ying-ying, Yu Zi-chao, Wang Qing-ming, Xu Tie-jun
    2011, 15 (49):  9169-9173.  doi: 10.3969/j.issn.1673-8225.2011.49.011
    Abstract ( 421 )   PDF (1450KB) ( 306 )   Save

    BACKGROUND: PNMDA receptors are major specific excitatory amino acid glutamate receptors in the brain. Studies have shown that NMDA receptors are involved in many important physiological and pathological processes of the central nervous system.
    OBJECTIVE: To determine the NMDA receptor-mediated whole-cell current changes during the development of neural stem cells (NSCs) and neural precursor cells (NPCs) in SD rat hippocampus in vitro.
    METHODS: NPCs were induced for 1, 3, 7 and 9 days. Nestin and βⅢ-tubulin positive cells were assessed by immunocytochemistry. Currents were recorded at -60 mV using patch clamp techniques. The NSCs and NPCs were divided into control and experimental groups. For the control group, cells were perfused with Mg2+-free extracellular solution. For the experimental group, cells were respectively perfused with 10, 20, 50 and 100 μmol/L NMDA.
    RESULTS AND COUCLUSION: There were no NMDA receptor-induced currents recorded in NSCs. No NMDA receptor-induced currents were detected in NPCs after 1-2 days of differentiation. Current records were obtained after 3 and 7 days of differentiation, and the currents were strengthened with the increasing dose of NMDA. The currents obtained after 7 days of differentiation were larger than those obtained after 3 days of differentiation at the same dose of NMDA. It was indicated that: ①NMDA-mediated currents in NSCs were not detected during experiments. ②NMDA-mediated currents in NPCs were detected after 3 days of differentiation. ③With the differentiation progress of NPCs, NMDA-mediated currents increased.

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    Differentiation of rat embryonic cerebral cortex neural stem cells cultured by erythropoietin in vitro
    Yuan Li-li, Du Hong-mei, Guan Ying-jun, Kong You-hua
    2011, 15 (49):  9174-9177.  doi: 10.3969/j.issn.1673-8225.2011.49.012
    Abstract ( 244 )   PDF (1056KB) ( 512 )   Save

    BACKGROUND: Erythropoietin (FPO) is early known as a kind of growth factor. In recent years, there are many studies about the protective effect of erythropoietin on the central nervous system in vivo.
    OBJECTIVE: To explore the effects of EPO on the apoptosis and differentiation of rat embryonic cerebral cortex neural stem cells in vitro.
    METHODS: The embryonic cerebral cortex were isolate from Sprague-Dawley rats that pregnant for 14 days under sterile conditions, the cells were cultured and differentiated in the suspension medium and then adherent induction. The neural stem cells were detected by nestin immune cell fluorescence staining, microtubule-associated protein 2 (MAP-2) and glia fibrillary acid protein was used to detect the differentiation of neural stem cells. The third passage of neural stem cells were obtained and 0.5, 5, 50, 500 U/mL EPO were added into the medium, and in the control group, there was no EPO adding. Caspase-3 was used to detect the apoptosis of neural stem cells, and MAP-2 was used to detect the differentiation of neural stem cells to neuron.
    RESULTS AND CONCLUSION: The expression of the caspase-3 in neural sphere was decreased after added EPO (≥5 U/mL), and the expression of the MAP-2 positive cells was increased obviously after differentiation (P < 0.01). The present results suggest that EPO can decrease the apoptosis of the neural stem cells and promote neural stem cells to differentiate into neurons in vitro.

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    Effects of engrafted neural stem cells derived from fetal rat on model rat with Parkinson's diseases
    Jiang Ming, Chen Xin-cheng, Wu Min, Deng Yin-sheng, Tao Yi
    2011, 15 (49):  9178-9181.  doi: 10.3969/j.issn.1673-8225.2011.49.013
    Abstract ( 335 )   PDF (615KB) ( 315 )   Save

    BACKGROUND: Stem cells transplantation is one of the potential methods for the treatment of Parkinson's diseases (PD).
    OBJECTIVE: To investigate the effect of neural stem cells engrafted into the striatum on rotational behavior and dopamine concentration of rat model with PD.
    METHODS: Rat models of PD were established by point injection with 6-hydroxy dopamine to damage nigrostriatal, the 1×106 (total 20 μL) passage 3 fetal rat neural stem cells or normal saline were injected into the striatum of the established rat model.   
    RESULTS AND CONCLUSION: After neural stem cells were transplanted into the striatum, the rotational behavior of the PD rat was improved obviously. Three weeks later, the results of immunohistochemistry detection showed that the number of tyrosine hydroxylase-positive cells in the substantia nigra of transfected PD rat was increased obviously, and the tyrosine hydroxylase-positive cells could be seen in the striatum. Fluorescence microscope observation showed that the Hoechst 33324d marked stem cells were more intensive near the site of transplantation and migrated to distant part. Eight weeks later, high performance liquid chromatography analysis showed that dopamine concentration in the striatum of stem cells transplanted PD rats was significantly higher (P < 0.01). It indicates that intracerebral neural stem cells transplantation can reduce the rat substantia nigra dopaminergic neuronal damage caused by 6-hydroxy dopamine, and improve the rotational behavior of PD rat.

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    Treatment of traumatic brain injury by human umbilical cord mesenchymal stem cells transplantation combined with C5a receptor antagonist
    Lü Jia-xi, Yin Hong
    2011, 15 (49):  9182-9185.  doi: 10.3969/j.issn.1673-8225.2011.49.014
    Abstract ( 252 )   PDF (549KB) ( 327 )   Save

    BACKGROUND: C5a is an important mediator and chemokine of inflammation, which plays an important role in the process of traumatic brain injury (TBI) pathogenesis.
    OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) transplantation and C5a receptor antagonist on TBI.
    METHODS: Sprague-Dawley rats were prepared to the heavy-duty hydraulic traumatic brain injury models and they were randomly divided into three groups: TBI group, hUC-MSCs transplantation group and hUC-MSCs transplantation plus C5a receptor antagonist group (C5a+hUC-MSCs group).
    RESULTS AND CONCLUSION: At 1-4 weeks after transplantation, the modified neurological severity scores (mNSS) were lower in the hUC-MSCs and C5a+hUC-MSCs groups than in the TBI group (P < 0.05, 0.01), while mNSS were decreased significantly in the C5a+hUC-MSCs group. The average time of escape latency was gradually decreased in each group; however, it was much shorter in the C5a+hUC-MSCs group than in the other two groups at 3-5 days (P < 0.05). The frequency of platform passing and the percentage of swimming distance traveled in the previous target quadrant in the C5a+hUC-MSCs group were significantly greater than those in the TBI group and hUC-MSCs group. It is indicated that UC-MSCs transplantation combined with C5a receptor antagonist can significantly improve the neurological function in the rats.

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    Proliferation and directed differentiation of neural stem cells and olfactory ensheathing cells after co-transplantation into the brain of Alzheimer’s disease rats
    Sheng Bao-ying, Li Yang, Jiang Yao-jia, Wei Chun-jie, Li Ying, Ren Xiu-min
    2011, 15 (49):  9186-9189.  doi: 10.3969/j.issn.1673-8225.2011.49.015
    Abstract ( 327 )   PDF (613KB) ( 377 )   Save

    BACKGROUND: The number of neurons is reduced in the brain of Alzheimer’s disease rats, and the proliferation and differentiation ability of transplanted neural stem cells (NSCs) into neurons is limited.
    OBJECTIVE: To investigate the effects of olfactory ensheathing cells (OECs) on the proliferation and differentiation of NSCs to cholinergic neurons after co-transplantation into the brain of Alzheimer’s disease rats.
    METHODS: OECs and NSCs were separately cultured in vitro, and the NSCs labeled with 5-bromodeoxyuridine (BrdU) before transplantation. Normal saline, NSCs and NSCs+OECs were separately transplanted into the hippocampi of Alzheimer’s rats. At 7, 14, 21 and 28 days after transplantation, the immunohistochemical staining was used to detects the expression of BrdU and choline acetyltransferase (ChAT) in the slice of rat brain tissues to analyze the transplanted cells’ proliferation and differentiation.
    RESULTS AND CONCLUSION: The condition of proliferation and differentiation of the NSCs co-transplanted with OECs was the best. The BrdU-positive cells and ChAT-positive cells of the OECs and NSCs co-transplantation group were the most of the three groups (P < 0.01). OECs can promote the proliferation and differentiation of NSCs to cholinergic neurons after co-transplantation in the brain of Alzheimer’s disease rats.

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    Bone marrow mesenchymal stem cells improve cardiomyocyte contractility in rats with heart failure
    Wang Tong, Zheng Shao-xin, Zhou Chang-qing, Weng Yin-lun, Wen Zhu-zhi
    2011, 15 (49):  9190-9193.  doi: 10.3969/j.issn.1673-8225.2011.49.016
    Abstract ( 274 )   PDF (1608KB) ( 310 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) transplantation can significantly improve cardiac function.
    OBJECTIVE: To investigate the effect of BMSCs on cardiomyocytes contractility and dilatation in rats with heart failure after myocardial infarction.
    METHODS: Left anterior descending coronary arteries of 10 Sprague Dawley rats were ligated in order to establish the myocardial ischemia model. The model rats were randomly divided into two groups: the experimental group and the control group. Four weeks later, 5×106/L PKH26 labeled BMSCs were injected into the infracted myocardium in the experimental group and in the control group, only injected with PBS.  
    RESULTS AND CONCLUSION: Compared with ligation, the cardiac function in two groups after modeling for 4 weeks was improved significantly (P < 0.01). After treated with BMSCs administration, the cardiac function in experimental group was improved significantly (P < 0.05). The amplitude of shortening/relengthening and contractile velocity of cardiomyocytes in experimental group was significantly increased (P < 0.01). The cardiomyocytes with contractility differentiated from BMSCs could be seen under fluorescence microscope. The injection of BMSCs could significantly increase the shortening/relengthening function of rat cardiomyocytes and was one of the important mechanisms in BMSCs improved of heart failure rats.

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    Transplantation of endothelial progenitor cells from human peripheral blood improves limb ischemia
    Wang Hui, Li Nan-lin, Yuan Shi-fang, Chen Jiang-hao, Li Yong-ping, Yuan Jun, Yi Jun, Wang Ling
    2011, 15 (49):  9194-9197.  doi: 10.3969/j.issn.1673-8225.2011.49.017
    Abstract ( 300 )   PDF (1398KB) ( 309 )   Save

    BACKGROUND: Transplantation of endothelial progenitor cells provides us a new treatment options on limbs ischemia which is hard to cure in clinical.
    OBJECTIVE: To estimate the effect of transplantation of endothelial progenitor cells (EPCs) derived from human peripheral blood on the improvement of limbs ischemia. 
    METHODS: The mononuclear cells derived from human peripheral blood were obtained by Ficoll density gradient centrifugation. The expressions of specific markers of EPCs were detected after cultured and amplified in vitro for 6 days, and the adherence cells marked by fluorochrome were multipoint injected into the nude mouse model with hind limb ischemia in order to evaluate the therapeutic effect.
    RESULTS AND CONCLUSION: The adherence cells derived from mononuclear cells could express CD133, CD34 and vascular endothelial growth factor 2 (VEGFR-2). It indicated that we could culture and obtain the EPCs from mononuclear cells of human peripheral blood in vitro under certain condition. The necrosis and capillary density of the ischemia limbs of nude mice were improved obviously after EPCs transplantation (P < 0.05). Red and flavo-green fluorescently-labeled EPCs were observed in the paraffin section of ischemia limbs muscle. The transplanted EPCs can incorporate into the local capillary networks and improve the ischemia of posterior limb of nude mice.

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    Transplantation of human amnion membrane mesenchymal stem cells via tail vein for the treatment of cerebral ischemia –reperfusion injury in rats
    Xu Ying, Ling Wei-hua, Lu Shi-qi, Fang Qi, Li Jun-gen
    2011, 15 (49):  9198-9201.  doi: 10.3969/j.issn.1673-8225.2011.49.018
    Abstract ( 375 )   PDF (1300KB) ( 338 )   Save

    BACKGROUND: Amniotic mesenchymal stem cells are an ideal seed cells sources for regenerative medicine.
    OBJECTIVE: To investigate the effect of human amnion membrane mesenchymal stem cells (hAMSCs) via tail vein for the treatment of cerebral ischemia injury.
    METHODS: The rat models of middle cerebral artery infarction were established, and underwent reperfusion at 2 hours after ischemia. 1.0×105 hAMSCs were transplanted through tail vein after 24 hours cerebral ischemia and the neurological severity score (NSS) was evaluated after 2 and 4 weeks cerebral ischemia. The rat without transplantation was as control.
    RESULTS AND CONCLUSION: After cerebral ischemia injury, the neurological score of rats was increased significantly; at 1 week after AMSCs transplantation, the neurological score of rats was decreased significantly (P < 0.05); at 2 and 4 weeks after AMSCs transplantation, the neurological score was further improved. Immunofluorescence staining showed that there were green fluorescence BrdU-positive cells in the rats cerebral tissue transplanted with hAMSCs, and the positive cells were mainly located around the ischemic area, subcortex and the lateral ventricle. It indicates that tail vein transplantation of hAMSCs can migrates to the ischemia area and improve the neurological function of rats.

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    Regulation of Wnt signaling pathway involved in the differentiation of hair follicle stem cells induced by lithium chloride
    Yang Bin, Xiang Meng-juan
    2011, 15 (49):  9202-9206.  doi: 10.3969/j.issn.1673-8225.2011.49.019
    Abstract ( 372 )   PDF (2023KB) ( 516 )   Save

    BACKGROUND: Previous studies showed that hair follicle stem cells (HFSCs) can undergo committed differentiation induced by lithium chloride (LiCl), but relating molecular regulation mechanism undisclosed.
    OBJECTIVE: To observe the changes of downstream molecules and target genes in Wnt signaling pathway during exogenous factor LiCl intervenes HFSCs and to investigate the role of LiCl in committed differentiation of HFSCs.
    METHODS: HFSCs were isolated from incised scalps of patients treated with rhytidectomy by two-steps enzyme digestion followed surgical microscope operation. HFSCs were subjected to antibodies-fluorescein isothiocy of cytokeratin 19 for identification. HFSCs was intervened by exogenous factor LiCl.
    RESULTS AND CONCLUSION: After 5 days HFSCs treated with LiCl, HFSCs volume got larger and the nucleus/cytoplasm ratio was reduced. In Wnt signaling pathway, the mRNA expression level of β-catenin and cyclinD1 increased, GSK3β and Tcf3 decreased, accompanied with c-myc increased at a lower concentration of LiCl and inversing decreased. LiCl can affect the expression level of β-catenin, GSK3β, Tcf3 ,c-myc and cyclinD1 involved in Wnt signal pathway, which may play an important role in HFSC differentiation.

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    Pten gene expression and phosphoproteomic analysis of the downstream of PI3K/Akt/mTOR signaling in human embryonic stem cells
    Zhou Rui-qing, Gong Yu-ping, Xing Hong-yun, Yang Xi
    2011, 15 (49):  9207-9210.  doi: 10.3969/j.issn.1673-8225.2011.49.020
    Abstract ( 386 )   PDF (1185KB) ( 954 )   Save

    BACKGROUND: Phosphoprteomic analyses of human embryonic stem cells (hESCs) in undifferentiated state and their differentiated derivatives have become a hotspot. The researchs suggest that phosphorylation events determine hESCs’ fate.
    OBJECTIVE: To investigate the expression of Pten mRNA and the key proteins of PTEN/Akt/mTOR signaling in human embryonic stem cells in order to provide the basic research for stem cells expansion and differentiation.
    METHODS: The hESCs were cocultured with the murine fetal fibroblasts to maintain undifferentiation and digested by collagenase for detection. The growth morphology of hESCs were observed, RT-PCR assay was used to determine Pten gene expression, and Western blot assay to detect the expression of p-PTEN, p-mTOR, p-P70S6K, p-4E-BP1 in hESCs.
    RESULTS AND CONCLUSION: The expression level of Pten was higher in hESCs and feeder cells than in K562, while the expression level of the key proteins in hESCs was lower than K562, especially p-4E-BP1 in hESCs. Over-activation of the PI3K/Akt/mTOR signaling may accelerate the proliferation of hESCs and deduce the apoptosis, and provide more cells for differentiation and regeneration medicine.

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    Effect of recombinant retrovirus pLXSN/basic fibroblast growth factor transfection on osteogenesis of human marrow stroma cells
    Zhou Rong-xing, Chen Hua, Guo Jun, Lü Hui-cheng, Yang Tie-yi
    2011, 15 (49):  9211-9214.  doi: 10.3969/j.issn.1673-8225.2011.49.021
    Abstract ( 292 )   PDF (1175KB) ( 321 )   Save

    BACKGROUND: In home and abroad have many studies on basic fibroblast growth factor (bFGF) transfection for improving the growth of muscle and vessel, but little studies on bFGF for improving the growth of osteogenesis.
    OBJECTIVE: To investigate the effect of recombinant retrovirus pLXSN/bFGF transfection on osteogenic potential of human marrow stroma cells (hMSCs). 
    METHODS: hMSCs isolated from health volunteers, cultured and purified in vitro were randomly divided into four groups. Retrovirus pLXSN/bFGF group with recombinant retrovirus pLXSN/bFGF was added into culture medium. Retrovirus pLXSN group with recombinant retrovirus pLXSN was added to take the place of retrovirus pLXSN/bFGF. Positive control group with dexamethasone, glycerophosphate and ascorbic were added into culture medium. Blank control group without any special treatment.
    RESULTS AND CONCLUSION: hMSCs were uniform spindle-shape fibroblast after several passages. After treatment, hMSCs in retrovirus pLXSN/bFGF and positive control groups gradually showed flat shape. Immunohistochemical staining showed that the level of bFGF expression in retrovirus pLXSN/bFGF group was higher than that of the other three groups. Mineralized nodules and bone collagen were formed, and alkaline phosphatase activity was improved in retrovirus pLXSN/bFGF and positive control groups. Recombinant of retrovirus pLXSN/bFGF gene transfection into hMSCs can promote the osteogenic potential of hMSCs.

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    Effect of scopion venom component Ⅲ on expression of toll like receptor 4 and p65 in lipopolysaccharide stimulated human leukemia monocytic THP-1 cells
    Song Xiang-feng, Niu Jie, Liu Yu-feng
    2011, 15 (49):  9215-9217.  doi: 10.3969/j.issn.1673-8225.2011.49.022
    Abstract ( 244 )   PDF (895KB) ( 344 )   Save

    BACKGROUND: In recent years, studies have shown that toll-like receptors and nuclear factor κB (NF-κB) have a close relation with the occurrence and development of leukemia. However, there were no reports about the effect of scorpion venom on toll-like receptors on the surface of tumor cells and NF-κB.
    OBJECTIVE: To investigate the effect of scopion venom component Ⅲ (SVC-Ⅲ) on expression of toll like receptor 4 (TLR4) and p65 in lipopolysaccharide (LPS) stimulated human leukemia monocytic THP-1 cells.
    METHODS: THP1 cells were inoculated in 24-well cells culture plates, and there were five groups: control group, LPS (1 mg/L) stimulation group and SVC-Ⅲ treatment group (LPS+ SVC-Ⅲ, 1, 10, 20 mg/L). Real-time PCR was employed to detect the expression of TLR4 and p65 mRNA in THP1 cells,Western blot was used to detected the expression of TLR4 and p65 protein.
    RESULTS AND CONCLUSION: The expression of TLR4 and p65 mRNA and protein of THP1 cells in LPS stimulation group was significantly higher than that in the control group (P < 0.01). However, the expression of TLR4 and p65 mRNA and protein of THP1 cells in SVC-Ⅲ treatment group was lower than that in LPS stimulation group (P < 0.01) and in dose-dependent manner. SVC-Ⅲ can down-regulate the expression of TLR4 and inhibit the activity of NF-κB in LPS stimulated THP1 cells in order to inhibit the cells proliferation.

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    Retinoic acid induces osteogenic differentiation of periodontal ligament stem cells from miniature swine in vitro
    Zhang Peng-tao, Zhong Liang-jun, Zhang Yuan, Zhang Yuan-ming, Xu Yan
    2011, 15 (49):  9218-9222.  doi: 10.3969/j.issn.1673-8225.2011.49.023
    Abstract ( 322 )   PDF (1334KB) ( 315 )   Save

    BACKGROUND: Recent studies have found that retinoic acid can induce the osteogenic differentiation of embryonic stem cells and multiple adult stem cells.
    OBJECTIVE: To observe the effect of retinoic acid on osteogenic differentiation of porcine periodontal ligament stem cells (PDLSCs).
    METHODS: Porcine PDLCs were harvested by using outgrowth method; PDLSCs were isolated by limited dilution of culture cells for single cell clone. Immunofluorescence was used to detect the expression of STRO-1 and immunocytochemistry to detect the expression of vimentin and pan-Cytokeratin (PCK) of porcine PDLSCs. Cell counting kit8 (CCK8) was applied to evaluated cell proliferation of PDLSCs. The colony formation of porcine PDLSCs ratio was assayed. Third passage PDLSCs were induced with mineralized conditional medium containing retinoic acid, ascorbic acid and β-glycerophosphate. Mineralized nodules were studied by Alizarin red S staining. Osteopontin, osteocalcin, collagen typeⅠ and collagen type Ⅲ were detected by immunocytochemistry.
    RESULTS AND CONCLUSION: Porcine PDLSCs expressed STRO-1 and vimentin, and the result of pan-Cytokeratin was negative. Porcine PDLSCs colony formation ratio was 2.8%. PDLSCs induced by retinoic acid showed positive expression of alkaline phosphatase at 14 days and positive expression of Alizarin red S at 21 days. Osteopontin, osteocalcin, collagen typeⅠ were positive but collagen type Ⅲ was negative at 21 days. These findings indicate that retinoic acid can be an effective inducer of osteogenic differentiation of porcine PDLSCs.

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    Differentiation of placenta mesenchymal stem cells to epidermal cells in vitro
    Wan Zhen-zhou, Feng Ya-song, Peng Hai-lin, Zhang Wei, Miao Zong-ning, Chen Ya-bao
    2011, 15 (49):  9223-9226.  doi: 10.3969/j.issn.1673-8225.2011.49.024
    Abstract ( 296 )   PDF (1154KB) ( 449 )   Save

    BACKGROUND: The pervious study has showed that placenta mesenchymal stem cells (PMSCs) can differentiate into bone, cartilage, and muscle and adipose mesenchymal stem cells.
    OBJECTIVE: To investigate the differentiation of PMSCs into epidermal cells in vitro.
    METHODS: PMSCs were isolated and cultured and the surface antigen was detected by flow cytometry.
    RESULTS AND CONCLUSION: Isolated PMSCs were positive for the markers CD105, CD29 and CD44 and negative for typical hematopoietic and endothelial markers CD34, CD45, CD106 and HLA-DR. The morphology of induced cells was obviously changed. Immunohistochemistry stain analysis showed positive for epidermal-specific markers like CK19 and CK10. The PMSCs have the capability to differentiate into epidermal cells in vitro. The placenta may prove to be a useful source of mesenchymal stem cells in skin tissue engineering and stem cells study.

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    Growth of actived keratocytes inhibited by limbal epithelial cells cultured in vitro
    Yu Xiao-fei, Xu Zhong-zhong, Du Lian-xin, Wang Li-ya
    2011, 15 (49):  9227-9230.  doi: 10.3969/j.issn.1673-8225.2011.49.025
    Abstract ( 311 )   PDF (1088KB) ( 265 )   Save

    BACKGROUND: When cornea injured, it can induce the phenotypic transition of keratocyte into the fibroblast, which causes the formation of the scar in the stroma and the loss of the vision.
    OBJECTIVE: To observe the interaction between the corneal epithelial cell in different regions and the actived keratocyte and to explore that whether the limbal epithelial cell can inhibit the growth of the actived keratocyte.
    METHODS: Enzyme digestion and mechanical dissociation were adopted to collect corneal epithelial cells and the keratocyte from central cornea, paramedian cornea, limbus, and then cultured in vitro. The changes of cell morphology and growth were recorded by phase contrast microscopy. The phenotype and proliferation capacity of the cultured cells were identified by immunofluorescent staining with antibodies for p63 and proliferating cell nuclear antigen (PCNA).
    RESULTS AND CONCLUSION: When the contact inhibition occured between corneal epithelial cells and fibroblasts, there was a define boundaries to divide them into two sections. In the limbal group, the positively expressing rates in the cultured corneal epithelial cells were higher for PCNA and p63, in the paramedian group the expression of PCNA is also higher and the expression of p63 was negative. In the central group, the expression of PCNA is lower and p63 was negative expression. Only the limbal group has a certain proportion of limbal epithelial stem cells. In limbal group the growth of fibroblasts was inhibited by limbal epithelial cells, after interaction four weeks, fibroblasts were degenerated into dead cell clusters. Because of the limbal stem cells deficiency in central group and paramedian group, the area of fibroblasts was increasing obviously, but the growth of corneal epithelial cells was inhibited and some were displaced by fibroblasts. It is indicated that limbal epithelial cells can inhibit the growth of actived keratocytes and degenerate them in vitro culture.

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    Expression of cancer-testis antigen in glioma stem cells
    Tang Jing-Feng, Xiao Shao-wen, Huang Yong-wang, Yang Bao-hua, Du Yi-qing
    2011, 15 (49):  9231-9234.  doi: 10.3969/j.issn.1673-8225.2011.49.026
    Abstract ( 266 )   PDF (871KB) ( 332 )   Save

    BACKGROUND: It is still unclear on the expression of cancer-testis antigen genes in glioma stem cells.
    OBJECTIVE: To detect the expression of cancer testis antigen genes in glioma stem cells.
    METHODS: Stem cells were cultured and isolated by neurosphere culture methods in DMEM/F12. Then the expression of cancer testis antigen genes were analyzed among parent cells, stem cells and differentiated cells by using semiquantitative RT-PCR and real time PCR.
    RESULTS AND CONCLUSION: We isolated tumor spheres from glioma cell lines by neurosphere culture. And then tumor spheres were cultured in medium containing serum, the morphology of the tumor spheres changed to adherent and they were not distinguishable from the parent cells. The expression of cancer testis antigen genes were analyzed by semiquantitative RT-PCR and quantitative RT-PCR and it was found that the expression were highest in stem cells compared with parent cells and differentiated cells. Cancer testis antigen genes may present as surface antigens in cancer stem cells and these findings suggest that cancer testis antigen genes may be helpful for targeted vaccine therapy against cancer stem cells in glioma patients.

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    Three-month therapeutic effect of bone marrow mesenchymal stem cells transplantation on the treatment of X-linked muscular distrophy mice
    Xu Yi-feng, Yang Xiao-feng, Wang Hong-mei, Zhang Yi-bin, Lü Nai-wu, Zhou Jin-xu
    2011, 15 (49):  9235-9238.  doi: 10.3969/j.issn.1673-8225.2011.49.027
    Abstract ( 347 )   PDF (1325KB) ( 305 )   Save

    BACKGROUND: Stem cells transplantation for treatment of X-limked muscular dystrophy (mdx) is the research hotspot. To compare with hematopoietic stem cells transplantation, mesenchymal stem cells transplantation has less risk.
    OBJECTIVE: To study the effect of bone marrow mesenchymal stem cells transplantation on the treatment of Duchenne muscular distrophy (DMD) mice.
    METHODS: Sixteen 4-week-old mdx mice were randomly divided into two groups: control group and transplanted group, eight mice in each group. C57BL/6 mice bone marrow mesenchymal stem cells or normal saline were injected intravenously and intramuscularly into the mice in transplanted group, respectively.
    RESULTS AND CONCLUSION: Three months after transplantation, the serum creatine kinase in transplanted group was decreased obviously, and the expression of dystrophin protein could be seen on the skeletal muscle membrane. There was on expression of dystrophin protein in control group. There were no obvious changes of motor function in two groups. Bone marrow mesenchymal stem cells transplantation had some therapeutic effect on the treatment of mdx mice, it might reduce the damage of muscle cell membrane and delay the progression of the disease.

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    Effects of Fasudil and RhoA silencing on the proliferation of Schwann cells
    Wang Yu-cheng, Zhou Jiang-bo
    2011, 15 (49):  9239-9243.  doi: 10.3969/j.issn.1673-8225.2011.49.028
    Abstract ( 291 )   PDF (777KB) ( 355 )   Save

    BACKGROUND: Schwann cells play an important role in peripheral nerve injury, regeneration and repair.
    OBJECTIVE: To determine whether Fasudil or RNAi-mediated RhoA gene silencing can optimize Schwann cells culture.
    METHODS: Schwann cells were divided into 5 groups: control group; 5 μmol/L Fasudil group; 10 μmol/L Fasudil group;
    15 μmol/L Fasudil group; 20 μmol/L Fasudil group; RhoA gene silencing group. Three days later, RT-PCR and Western blot were used to assess the expression of RhoA mRNA and RhoA protein. Cellular proliferation was determined by the cell growth curve
    (5 days) and MTT assay. The cell cycle was analyzed by flow cytometry.
    RESULTS AND CONCLUSION: There was no difference between 20 μmol/L and 15 μmol/L Fasudil groups (P > 0.05). Fasudil and RNAi-mediated RhoA gene silencing can promote the proliferation of Schwann cells, and 15 μmol/L Fasudil and RhoA gene silencing have the best effect.

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    Construction and identification of lentiviral vector encoding survivin gene of mice
    Yang Ping, Gou Xin, Zhou Qing-song, He Wei-yang, Yu Kun
    2011, 15 (49):  9244-9248.  doi: 10.3969/j.issn.1673-8225.2011.49.029
    Abstract ( 346 )   PDF (597KB) ( 323 )   Save

    BACKGROUND: The limited viability of mesenchymal stem cells restricts its application in treatment in vivo. Therefore, it is very important to find an experiment method to prolong the viability of mesenchymal stem cells so as to lay foundation for experiment in vivo.
    OBJECTIVE: To construct the lentiviral vector encoding survivin gene of mice with clone vector pLV.Des3d.P/hygro.
    METHODS: Oligo nucleic acid software was used to design a couple of primers according to survivin gene sequence of mice which is publicated on Genbank. The attB recombination site was added to the two ends of the couple of primers, and mSurvivin and IRES/EmGFP were amplified by PCR. The PCR product after BP reaction was transferred to Stb13, and the correct pDown-mSurvivin and pTail-IRES/EmGFP were selected and sequenced. The mixture of pDown-mSurvivin, pTail-IRES/EmGFP and pLV.Des3d.P/hygro was used to do LR reaction. The lentiviral vector pLV.EX3d.P/hygro-EF1A>mSurvivin>IRES/EmGFP was amplified and sequenced again.
    RESULTS AND CONCLUSION: The results of PCR and sequencing showed that the lentiviral vector encoding survivin gene of mice was constructed successfully, which lays foundation for the further study of survivin anti-apoptosis in mesenchymal stem cells.

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    Effect of bone marrow mesenchymal stem cells combined with various scaffold materials:A systematic review
    Fang Zhi-hui, Tan Jin-hai, Zeng Xian-tao,Yang Yong-bo, Li Sheng
    2011, 15 (49):  9249-9253.  doi: 10.3969/j.issn.1673-8225.2011.49.030
    Abstract ( 295 )   PDF (510KB) ( 408 )   Save

    BACKGROUND: Now, bone marrow mesenchymal stem cells (BMSCs) combined with different biological scaffold materials to composite repair bone defect, but the effectiveness is lack of a systematic review.
    OBJECTIVE: To explore the effect of BMSCs combined with different biological bone scaffold materials on repairing bone defect.
    METHODS: A computer based online search in CNKI, CBM, VIP and Wanfang to collect the published articles for the randomized controlled trails which using BMSCs as a seed stem cells composite with various scaffold materials from January 1990 to April 2011. Hand searching was also done to obtain any further information about the studies. The quality of the included studies was critically evaluated and data were analyzed.
    RESULTS AND CONCLUSION: The results showed that the BMSCs could effective compound with various biological materials, including gelatin sponge, poly lactic acid-glycolic acid copolymer porous materials, hydroxyapatite, calcium phosphate, coral, alginate and chitosan poly lactic acid, calcium sulfate, algal acid calcium, L-polylactide and platelet-rich plasma, and could differentiate to osteoblast. It is indicated that BMSCs can promote the ability of various support materials to repair new bone osteogenesis of bone defect.

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    Analysis of related policy, funds and outputs on stem cells in China
    Fu Jun-ying, Zhao Yun-hua
    2011, 15 (49):  9256-9261.  doi: 10.3969/j.issn.1673-8225.2011.49.032
    Abstract ( 318 )   PDF (778KB) ( 480 )   Save

    BACKGROUND: There are loose political and social environments for research and application on stem cell in China compared with in American and European countries, and Chinese governments have always paid many attentions to development of stem cells.
    OBJECTIVE: To know more about stem cell related policies released by Chinese governments, funds supported by the National Natural Science Foundation of China (NSFC), outcomes of publications and patents achieved, and the leading academies and companies in China.   
    METHODS: SCI database and Derwent database (http://isiknowledge.com), projects database funded by NSFC (http://159.226.244.28/portal/proj_search.asp), Wanfang database (http://www.wanfangdata.com.cn), VIP database (http://www.vmis.net.cn/yixue/index.asp), were searched by the first author. Search terms were “stem cell, stem cells” in English and Chinese. Thirty suitable papers related to policies and research developments on stem cell in China, 1 871 projects released by NSFC, 6 802 SCI papers published by Chinese authors, 1 225 patents applied to China Patent & Trademark Office were retrieved. 
    RESULTS AND CONCLUSION: Chinese governments continuously increase yearly investments, and have achieved huge outcomes. Number of SCI papers published by China lists the eighth all over the world, although there is a long distance from US. Chinese Academy of Science has released most of the related publications, and NSFC is the major source of funds. Number of stem cell related patents applied in China lists the seventh worldwide. The leading units applying patents are Zhejiang University, Academy of Military Medical Science and Shanghai University. China should promote stem cell research on the basic level, and strengthen its application research.

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    Clinical possibility of induction and differentiation of thyroid cells from stem cells
    Liao Hong-Qun
    2011, 15 (49):  9262-9266.  doi: 10.3969/j.issn.1673-8225.2011.49.033
    Abstract ( 396 )   PDF (682KB) ( 932 )   Save

    BACKGROUND: Studies about stem cell differentiation into thyroid cells are few. Research on embryonic stem cell differentiation into thyroid cell has made some achievements, but other types of stem cells have different features of differentiation, and whether they are able or unable to differentiate into thyroid cells remains to be further studied.
    OBJECTIVE: To overview the advantages and disadvantages as well as progress in the research of induction and differentiation of stem cells into thyroid cells.
    METHODS: A computer retrieval of PubMed database (1982-01/2011-04), Wanfang database (1982-01/2011-04), VIP database (1989-01/2011-04), and CNKI database (1994-01/2011-04) was done using the keywords of "stem cell, thyroid cells, induction, differentiation" in English and Chinese. A total of 1052 articles were retrieved, and finially 50 articles were included in result analysis.
    RESULTS AND CONCLUSION: Patients with irreversible hypothyroidism require lifelong levo-thyroxin (L-T4) replacement therapy that makes them feel discomfortable. In recent years, stem cell research is expected to solve these medical problems. Directed differentiation of embryonic stem cells may towards thyroid cells in vitro, but reject reaction and ethical issues limit the application of embryonic stem cells. Other types of stem cells have different features, they under the different induced condition can differentiate into cells of different lineage, and they have been widely used to treat various diseases. However, there are no reports about differentiation into thyroid cells.

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    Mesenchymal stem cells in the treatment of rheumatoid arthritis
    Chen Dong-xu, Bo Zhan-dong, Zhang Yi
    2011, 15 (49):  9267-9270.  doi: 10.3969/j.issn.1673-8225.2011.49.034
    Abstract ( 312 )   PDF (650KB) ( 477 )   Save

    BACKGROUND: Studies have demonstrated that bone marrow or adipose tissue derived mesenchymal stem cells have immunosuppressive and anti-inflammatory properties to have a broad application prospect in the treatment of rheumatoid arthritis and other autoimmune diseases.
    OBJECTIVE: To focus on the biological features of mesenchymal stem cells, especially immunosuppressive and anti-inflammatory properties, and to review the progress of mesenchymal stem cells in rheumatoid arthritis.
    METHODS: Foreign Medical Journal Full-Text Service and China National Knowledge Infrastructure databases were searched for articles concerning rheumatoid arthritis, culture and application of mesenchymal stem cells to rheumatoid arthritis published from 2005 to 2011. The key words were “marrow stem cells, rheumatoid arthritis, immunosuppressive” in Chinese and English. Repetitive articles were excluded. Finally, 24 articles were further summarized.
    RESULTS AND CONCLUSION: Mesenchymal stem cells appear as major regulatory cells in skeletal tissue homeostasis, controlling inflammation, immune response, fibrosis and tissue regeneration, but the efficacy and security of mesenchymal stem cells in the treatment of rheumatoid arthritis still need further studies.

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    Application progress of bone marrow mesenchymal stem cells in tissue-engineering research
    Zhao Da-cheng, Wang Yu-liang, Dang Yue-xiu, Zhao Lin
    2011, 15 (49):  9271-9274.  doi: 10.3969/j.issn.1673-8225.2011.49.035
    Abstract ( 268 )   PDF (641KB) ( 347 )   Save

    BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) play a significant role in tissue engineering as good seed cells.
    OBJECTIVE: To descript the experimental status of BMSCs in the bone, cartilage, liver, tendons and heart tissue engineering applications, clarify the momentousness role of BMSCs in tissue engineering.
    METHODS: A computer-based online search was conducted in NCBI database and PubMed database for literatures about the application of BMSCs in tissue engineering published from January 2000 to October 2010 with the key words of “Bone marrow mesenchymal stem cells, Mesenchymal stem cells, tissue engineering” in English. Exclusive criteria included repetitive studies and no original research.
    RESULTS AND CONCLUSION: According to the included and excluded criterion, a total of 33 references were selected to analysis. BMSCs proliferation activity, had potentiality of multi-directional differentiation, it could differentiated to osteoblasts, chondrocytes, hepatocytes, tendon cells and myocardial cells. Therefore, BMSCs might be favorable seed cell to utilize in tissue engineering, and treated clinical correlated diseases. This article confirmed BMSCs had a highly utilization value in tissue engineering, and was hoped to solve puzzlers problems applied with tissue engineering in the treatment of related diseases.

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    Endogenous neural stem cells for treatment of spinal cord injury
    Zhang Cheng, Chen Gang
    2011, 15 (49):  9275-9278.  doi: 10.3969/j.issn.1673-8225.2011.49.036
    Abstract ( 248 )   PDF (635KB) ( 297 )   Save

    BACKGROUND: Spinal cord injury is a serious neurological disease, and the life quality of spinal cord injury patients significantly declines. With the development of medical science, endogenous neural stem cells as a treatment strategy for spinal cord injury have attracted widespread attention in recent years.
    OBJECTIVE: To summarize the activation and differentiation of endogenous neural stem cells, as well as the mechanism in treating spinal cord injury.
    METHODS: A computer-based online search of Wanfang database and PubMed database from January 2002 to March 2011 was performed for related articles about neural stem cells for treatment of spinal cord injury, with key words of “endogenous neural stem cells, spinal cord injury, differentiation” in Chinese and in English, respectively, by screening titles and abstracts. The documents associated with endogenous neural stem cells for treatment of spinal cord injury were selected, and those published recently or in authoritative journals were preferred in the same field. Totally 340 articles were obtained in initial retrieval, and 20 articles of them were chosen based on inclusion criteria.
    RESULTS AND CONCLUSION: Endogenous neural stem cells have unique advantages as a new treatment. They can not only avoid the immune rejection and ethical issues, but also exist in adult individuals. Most endogenous neural stem cells can differentiate into astrocytes and oligodendrocytes rather than neurons. More neural stem cells can differentiate into neurons through the regulation of signal transduction pathways, which plays an important role in the treatment of spinal cord injury.

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    Differentiation of bone marrow mesenchymal stem cells into endothelial cells and clinical application
    Kang Shao-ping, Jiang Wen-jing, Li Ping
    2011, 15 (49):  9279-9282.  doi: 10.3969/j.issn.1673-8225.2011.49.037
    Abstract ( 429 )   PDF (527KB) ( 341 )   Save

    BACKGROUND: Studies have preliminarily verified that the bone marrow mesenchymal stem cells (BMSCs) can differentiate into endothelial cells.
    OBJECTIVE: To review the differentiation of BMSCs into endothelial cells and the progress of clinical research.
    METHODS: PubMed database was searched for articles published from 2000 to 2011 with the key words of “bone marrow mesenchymal stem cell, endothelial cell, differentiation, tissue engineered. The language was limited in English. Meanwhile, Wanfang database was searched for articles published from 2000 to 2011 with the key words of “bone marrow mesenchymal stem cell, endothelial cell, differentiation, clinical application” in Chinese.Totally 418 articles were retrieved, and finally 31 articles were included. .
    RESULTS AND CONCLUSION: BMSCs have the potential of self renewal and multiplex differentiation. In certain conditions, they can be induced into endothelial cells. Endothelial cells are the important components of tissue-engineered blood vessels. BMSCs for endothelial construction can promote the regeneration of blood vessels in tissue-engineered organs, provide blood supply and also lay the foundation for the treatment of ischemic diseases in clinic.

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    Influence of Chinese medicines on mesenchymal stem cells culture and differentiation as well as stem cells transplantation for cardio-cerebrovascular diseases
    Shi Jing-yang, Tang Xue-min
    2011, 15 (49):  9283-9286.  doi: 10.3969/j.issn.1673-8225.2011.49.038
    Abstract ( 262 )   PDF (598KB) ( 349 )   Save

    BACKGROUND: Chinese medicine and compound recipe can promote mesenchymal stem cells proliferation and differentiation. Under certain condition, MSCs can differentiate into cardiomyocyte-like cells and neural-like cells and improve cardio-cerebrovascular disease.
    OBJECTIVE: To discuss influence of Chinese medicines on different source-derived stem cells and analyze application of stem cells in cardio-cerebrovascular diseases.
    METHODS: A computer-based online search of Wanfang and VIP database was performed for related articles published between 1990 and 2011 with key words “Chinese medicine, stem cell, proliferation, differentiation, target” in Chinese. Articles regarding roles of Chinese medicines in stem cells proliferation and differentiation, transplantation of stem cells for brain ischemia or brain injury in combination with Chinese medicines, and Chinese medicine-treated stem cells for cardiovascular diseases, were included. After reading the titles and abstracts, 30 articles were analyzed.
    RESULTS AND CONCLUSION: Chinese medicines can directly target to stem cells in vivo to promote their proliferation and differentiation and can change microenvironment of stem cells to promote cells survival and functional building or regulate immune function to relieve or eliminate rejection. Although stem cells transplantation has not frequently used in combination with Chinese medicines for cardio-cerebrovascular diseases, they have exhibited special clinical values. Therefore, transplantation of stem cells in combination with traditional Chinese medical theory may provide novel approaches for stem cells transplantation and effective methods for application of Chinese medicines in life science domains.  

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    New development of regenerative medicine: DNA methylation and cell reprogramming
    Chen Li-li, Cui Yu-xia, Yang Shui-xiang
    2011, 15 (49):  9287-9290.  doi: 10.3969/j.issn.1673-8225.2011.49.039
    Abstract ( 265 )   PDF (453KB) ( 430 )   Save

    BACKGROUND: A small number of growth factors are sufficient to reprogram somatic cells into pluripotent stem cells.
    OBJECTIVE: To explore the epigenetic mechanism in cellular reprogramming and regulating, and to reveal epigenetic alterations, the stability of epigenetic modifications and their effects on genome expression.
    METHODS: Pubmed database and CNKI database from 1990 to 2008 were retrieved by computer with the key words of “DNA methylation; cell reprogramming; stem cells”.
    RESULTS AND CONCLUSION: Though cellular differentiation is unidirectional and irreversible, this process can be changed by reprogramming. Some growth factors are sufficient to reprogram somatic cells into pluripotent stem cells. So far, the introduction of transcription factors, DNA demethylation and the apparent genetic modification have been used to induce reprogramming. By understanding the molecular mechanism, the apparent changes in genetic regulation, the stability of epigenetic marks and their effects on expression of the genome are significant in the development of gene therapy. However, many issues still need further studies, such as: Is AID or DNA demethylation enzyme plays an important role in DNA demethylation? What are the requirements for DNA demethylation? Can demethylation take place in CpG fragments of tumor cells? Are cancer cells more difficult to re-programming?

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    Stem cells transplantation for treatment of diabetes mellitus
    Li Xue, Chen Qiu
    2011, 15 (49):  9291-9294.  doi: 10.3969/j.issn.1673-8225.2011.49.040
    Abstract ( 290 )   PDF (574KB) ( 425 )   Save

    BACKGROUND: Stem cells transplantation can treat diabetes mellitus on cell and gene levels to achieve clinical cure.
    OBJECTIVE: To review the research progress addressing stem cells transplantation for diabetes mellitus.
    METHODS: A computer based search was conducted in PubMed database and Chinese Journal Full-text database for relevant articles published from January 2000 to June 2010, with keywords of “stem cells, diabetes mellitus, transplantation, insulin-secreting cell, treatment” in English and Chinese, respectively. A total of 27 articles were reviewed.
    RESULTS AND CONCLUSION: Diabetes mellitus is a group of metabolic disease characterized by chronic high blood sugar, which is caused by absolute or relative deficiency of insulin. Currently there is no treatment to cure it. Stem cells have a strong self-renewal and differentiation potential, which can be induced to differentiate into insulin-secreting cells to regulate blood sugar in vivo and in vitro, and they become the new resource of a substitute of islet β cells. At present, stem cells transplantation on treatment of diabetes mellitus has made some progress in animal experiments and small-scale clinical studies.

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    Mature epidermal cells dedifferentiate towards epidermal stem cells
    Zhao Xue-kai, Liang Zi-qian, Zhang Xian-fa, Zong Shou-kai
    2011, 15 (49):  9295-9298.  doi: 10.3969/j.issn.1673-8225.2011.49.041
    Abstract ( 314 )   PDF (556KB) ( 349 )   Save

    BACKGROUND: Epidermal stem cells as seed cells have been paid great attention on the research of wound repair and regeneration with its specific biological advantages with the development of molecular biology and bioengineering.
    OBJECTIVE: To review the recent advances in the mature epidermal cells dedifferentiating towards epidermal stem cells.
    METHODS: The first author retrieved PubMed database, Highwire database (Highwire.standford.edu) and CNKI for articles published from January 1998 to January 2011. The key words were “dedifferentiation, epidermal stem cell, epidermal cell, induction” in English and Chinese. A total of 2 754 literatures were collected, and finally 46 papers were included in result analysis.
    RESULTS AND CONCLUSION: The dedifferentiated epidermal stem cells have some characteristics of native epidermal stem cells and perhaps they may act as a new vehicle for wound repair and regeneration. Both of the native epithelial stem cells and the dedifferentiated epithelial stem cells can promote the wound repair and their effects were on the same level. Because differentiation from epidermal cells into epidermal stem cells has many questions, so far, there are many animal researches, but its extensive clinical application deserves further development.

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    Research advances in cardiac stem cells
    Yang Wen-ling, Zhao Xiao-hui
    2011, 15 (49):  9299-9302.  doi: 10.3969/j.issn.1673-8225.2011.49.042
    Abstract ( 337 )   PDF (280KB) ( 351 )   Save

    BACKGROUND: A large amount of studies have demonstrated that stem cells in the heart of mammal participate in heart self-renewal and endogenous repair.
    OBJECTIVE: To summarize the source, classification, features of cardiac stem cells and application in heart disease.
    METHODS: A computer-based online search of PubMed database was performed for articles published between January 2000 and December 2010 related to the source, classification, features of cardiac stem cells and its effects on myocardial regeneration with key words “cardiac stem cell”. Clinical studies and basic studies were all included. Repetitive studies and Meta analysis were excluded. Finally, 32 articles were included.
    RESULTS AND CONCLUSION: Cardiac stem cell is a type of stem cells in the heart, with properties of self-renewal and cloning proliferation. It can differentiate into cardiomyocyte and endothelial cell and plays a role in heart injury repair to improve heart function. Cardiac stem cells have been in vitro isolated, cultured and amplified to transplant into animal heart, which lays a foundation for clinical application. However, some questions remain poorly understood as to the homeostasis and dynamic changes, processes involved in their differentiation into heart functioning cells, as well as influential factors and regulation.

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    Ethical and psychologial issues of embryonic stem cell researches
    Dong Xiao-xin, Ren Xiao-min, Dong Ya-ni
    2011, 15 (49):  9303-9306.  doi: 10.3969/j.issn.1673-8225.2011.49.043
    Abstract ( 260 )   PDF (543KB) ( 414 )   Save

    BACKGROUND: Embryonic stem cell research has important medical value and is widely used in fields of gene therapy, tissue engineering and pharmacy, these research achievements can provide a new hope for treatment of diseases, however, causes many ethical and psychological issues.
    OBJECTIVE: To summarize the ethical and psychological issues in current studies regarding the embryonic stem cells.
    METHODS: A computer-based retrieval was performed in PubMed database, VIP database and Wanfang database from January 2001 to December 2010, for documents regarding the ethical and psychological issues in stem cell researches. The key words were "embryonic stem cells, morality, psychological" in English, and "embryonic stem cells, ethics, psychology" in Chinese. Totally 122 papers were screened out and 33 of them were ultimately included in the analysis.
    RESULTS AND CONCLUSION: The ethical issues of embryonic stem cell researches include the source of stem cells, stem cell cloning, stem cells and life, the existing problems of stem cell research and the related ethical specifications. The psychological issues of stem cell research mainly focus on the understanding of stem cell research, especially cloning technology. A number of researches are involved in the psychological changes of patients after hematopoietic stem cells transplantation. We have made a breakthrough in field of embryonic stem cell research, thus bring a revolutionary impact on the medical and health issues.

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    Non-myeloablative allogeneic hematopoietic stem cells transplantation for the treatment of elderly patients with acute leukemia
    Lu Jing-yuan, Lu Quan-yi
    2011, 15 (49):  9307-9310.  doi: 10.3969/j.issn.1673-8225.2011.49.044
    Abstract ( 282 )   PDF (660KB) ( 334 )   Save

    BACKGROUND: Non-myeloablative allogeneic hematopoietic stem cells transplantation can reduce the intensity of conditioning regimen, and decrease the transplantation related mortality rate.
    OBJECTIVE: To analyze the efficacy and safety of non-myeloablative allogeneic hematopoietic stem cells transplantation for the treatment of elderly patients with acute leukemia.
    METHODS: A 71-year-old female patient with acute mononuclear cell leukemia M5b underwent the non-myeloablative allogeneic hematopoietic stem cells transplantation. The donor was the younger brother of the patient who was 60 years old, ABO blood type was the same and the HLA loci were mismatched. The conditioning regimen contained fludarabine, thiotepa, cyclophosphamide and Myleran, human anti-thymocyte globulin(ATG), basiliximab, cyclosporine, mycophenolate mofetil and gamma globulin were used for graft-versus-host disease (GVHD) prophylaxis. The prophylaxis and support therapy of complication after transplantation consisted of Ganciclovir for cytomegalovirus infection, prostaglandin E1, salvia Miltiorrhiza; Heparin and ursodeoxycholic acid for veno-occlusive disease; Granulocyte colony-stimulating factor (G-CSF) 5 μg/kg per day was used when the absolute neutrophil count decreased to 0.5×109 /L. Red cells and platelet were infused if necessary. Peripheral blood stem cells (PBSC) of the donor were mobilized with G-CSF for 7 successive days. PBSC were collected and infused to the patient at the 6th and 7th days.
    RESULTS AND CONCLUSION: Hemopoiesis and immunology reconstruction was completed at the 28th day after transplantation. Results of short tandem repeat (STR) detection shows that the DNA from the donor was the same as the patient at the 50th day after transplantation. The patient suffered from hemorrhagic cystitis and sepsis (Burkholderiacepacia) after transplantation for 20-30 days. Two months later, the patient presented with jaundice and hepatic impairment and she was diagnosed as having acute GVHD (liver, grade 3). These were controlled with antibiotics and adjusting the regime of immunosuppressive drugs. These findings indicate that non-myeloablative allogeneic hematopoietic stem cells transplantation based on a pretreatment regime containing fludarabine is safe and effective for elderly acute leukemia patients with a variety of complications.

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