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03 September 2011, Volume 15 Issue 36 Previous Issue    Next Issue

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CXCL12/CXCR4 biology axis effects on the repair of spinal cord injury with bone marrow mesenchymal stem cells Fan Dong-yan, Liu Yan, Xu Fu-chun, Wang Ping 2011, 15 (36):  6651-6656.  doi: 10.3969/j.issn.1673-8225.2011.36.001 Abstract ( 323 )   PDF (1857KB) ( 399 )   Save BACKGROUND: The research discovered that bone marrow mesenchymal stem cells (BMSCs) through different approaches can migrate to the injured site of the spinal cord, and play curative effects. OBJECTIVE: To discuss the role of CXCL12/CXCR4 biology axis on the migration of BMSCs into the injured site after spinal cord injury (SCI). METHODS: SCI models were prepared using modified spinal damage method. There was no intervention in the sham operation group beside skin open. In the model group, lumbar intrathecal injection of 5 μL normal saline was used at 2 days after modeling; in the transplantation group, lumbar intrathecal injection of 5μL BMSCs was administrated at 2 days after modeling. RESULTS AND CONCLUSION: Under the fluorescence microscope, a large mass of labeled cells gathered at the injured site, however, only a few of labeled BMSCs could be seen 1 cm distant from the distal injured site. BMSCs expressed the medium level of CXCL12, and CXCR4 also had a low level expression in the BMSCs. Seven days after SCI, partial CXCL12 expression strengthened, mainly in the cortex region of SCI, but there were no massive expressions of CXCL12 1 cm outside the injured site. CXCR4 protein expression did not present with a time-effect manner. CXCR4 transcriptional level in the transplantation group was obviously higher than that in the sham operation group and model group. At 14 days after SCI, CXCL12 transcriptional level reached the peak, and lowered at 21 days. The local CXCL12 transcriptional level was remarkably higher than that at the distal end. CXCR4 also expressed at the injured site in a time-independent manner. The partial CXCR4 transcriptional level was slightly higher than that at the distal end, but there was no significant difference. The results indicated that the CXCL12/CXCR4 biology axis participates in the migration of BMSCs into the injured zone after SCI. Related Articles | Metrics

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Effect of DNA demethylation on telomerase reverse transcriptase expression in rat bone marrow mesenchymal stem cells Zhao Hong-xian, Lu Wei, Guo Yong, Chen Xia 2011, 15 (36):  6657-6660.  doi: 10.3969/j.issn.1673-8225.2011.36.002 Abstract ( 240 )   PDF (1214KB) ( 238 )   Save BACKGROUND: DNA demethylation is an important epigenetic modification, which plays a vital role in telomerase regualtion of tumor cells. It is not clear to effect of DNA demethylation on telomerase activity in bone marrow mesenchymal stem cells. OBJECTIVE: To observe the effect of DNA demethylation on proliferation and telomerase reverse transcriptase expression in bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were isolated by using the method of adhesive culture of whole bone marrow. In the third passage bone marrow mesenchymal stem cells, 5-azacytidine was added to 0, 3, 6, 12, 24 μmol/L. CD44, CD45 and telomerase reverse transcriptase were dectcted by immunocytochemistry, and effect of DNA demethylation on proliferation was detected by MTT method at 1, 2, 3, 5, 7 days after 5-azacytidine intervention. RESULTS AND CONCLUSION: Compared with the control group, after 24-hour intervention of 5-azacytidine, 5-azacytidine with 3, 6, 12, 24 μmol/L increased significantly cell proliferation without dose-dependence (P < 0.05). After 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased significantly cell proliferation without dose-dependence (P  < 0.05). After 72-hour intervention of 5-azacytidine, 5-azacytidine with 12, 24 μmol/L decreased significantly cell proliferation without dose-dependence (P < 0.05). After 5-day and 7-day intervention of 5-azacytidine, 5-azacytidine has no effect on cell proliferation (P > 0.05). Compared with the control group, after 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased significantly telomerase reverse transcriptase expression in bone marrow mesenchymal stem cells without dose-dependence (P  < 0.05). In certain concerntration and time, 5-azacytidine can increase proliferation and expression of telomerase reverse transcriptase in bone marrow mesenchymal stem cells. Related Articles | Metrics

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Mesenchymal stem cells can differentiate to epidermal cells and express keratins in vitro  Li Ling-yun, Du Rong, Fan Xi-ming 2011, 15 (36):  6661-6664.  doi: 10.3969/j.issn.1673-8225.2011.36.003 Abstract ( 345 )   PDF (1054KB) ( 296 )   Save BACKGROUND: Animal experiments have proved that bone marrow mesenchymal stem cells (MSCs) can mostly differentiate into epidermal cells.    OBJECTIVE: To investgate the feasibility of inducing MSCs into epidermal cells and the expression of keratins. METHODS: MSCs were extracted by Ficoll-Paque density gradient centrifugation. The expressions of CD34 and CD44 in MSCs cultured for three passages were deteced by immunocytochemical staining and flow cytometric examinations. The third passage MSCs were culured in 30% conditioned medium and induced to differentiate into epidermal cells. The changes, of cytokeratin 19 were detect by immunocytochemical staining and the morphological features were observed after being induced. RESULTS AND CONCLUSION: The MSCs extracted by Ficoll-Paque density gradient centrifugation were pure. After being induced with conditioned medium, cytokeratin 19 in MSCs were observed, indicating that MSCs may be induced into epidermal cells． Related Articles | Metrics

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Notch1 signaling system regulates the differentiation of bone marrow mesenchymal stem cells into cardiomyocytes Niu Ping, Zhao Yue-qiang, Huang Xing-yuan 2011, 15 (36):  6665-6668.  doi: 10.3969/j.issn.1673-8225.2011.36.004 Abstract ( 219 )   PDF (1068KB) ( 260 )   Save BACKGROUND: Notch signaling system has an important role of regulating the differentiation of bone marrow mesenchymal stem cells (BMSCs). However, there are no reports about Notch signaling system effect on differentiation and mechanism of BMSCs into cardiomyocytes. OBJECTIVE: To investigate the regulation of differentiation of BMSCs into cardiomyocytes by Notch signaling system. METHODS: BMSCs were cocultured with neonatal rat ventricular myocytes in vitro. Jagged1, an activator of the Notch signaling system, was added into the culture medium of the experiment group. The control group was added with Jagged1 and DAPT, an inhibitor of the Notch signaling system. The cells were cultured in the medium added with phosphate-buffered saline used as blank control group. Then the cultured cells were collected. The expression of Notch signal and TnT was detected by RT-PCR and immunohistochemistry. RESULTS AND CONCLUSION: BMSCs can differentiate into cardiomyocytes in vitro. Compared with the two control groups, the differentiate percentage was higher in the experimental group. RT-PCR showed that the expression level of the receptor Notch1 and the ligand Jagged1 was significantly higher. Immunohistochemistry showed a significantly increased expression of TnT. When the Notch signaling system is activated, the differentiation of BMSCs into cardiomyocytes enhances, while the effect can be inhibited by DAPT. Related Articles | Metrics

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Induced differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro Zhang Tao, Zhao Zhen-guo, Gao Hui 2011, 15 (36):  6669-6672.  doi: 10.3969/j.issn.1673-8225.2011.36.005 Abstract ( 260 )   PDF (1142KB) ( 293 )   Save BACKGROUND: The study of the induction of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells majorly concentrates on the induction of different inducing factors, and pays little attention on the induction of micro environment. OBJECTIVE: To investigate the differentiation of rat BMSCs into hepatocyte-like cells. METHODS: The rats BMSCs were isolated by density gradient centrifugation and purified by adherence cultivation, morphology feature of BMSCs were observed by microscope. The fetal liver cells were isolated by adherence cultivation after digesting the rats embryo liver of 3 weeks with collagenase. The negative control group was cultured in L-DMEM containing 10% fetal bovine serum. The purified BMSCs in the induced group were cultured in L-DMEM containing 10% fetal bovine serum with hepatocyte growth factor (HGF) and fetal liver cells. RESULTS AND CONCLUSION: Compared with those in non-induced BMSCs, the levels of alpha-fetoprotein and albumin in the induced-BMSCs were higher (P < 0.01). Both glycogen and CK-18 were positive in the induced BMSCs. BMSCs can differentiate into hepatocyte-like cells with hepatic phenotype and function in the presence of HGF and fetal liver cells, which may be used as a kind of cell resources to treat severe hepatic disease. Related Articles | Metrics

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Osteogenous differentiation of bone marrow mesenchymal stem cells transfected with bone morphogenetic protein 4 and bone morphogenetic proteins 4/7 Yuan Shao-hui, Liu Wei, Wu Bin-qi, Han Xi-guang, Bi Zheng-gang 2011, 15 (36):  6673-6678.  doi: 10.3969/j.issn.1673-8225.2011.36.006 Abstract ( 255 )   PDF (690KB) ( 397 )   Save BACKGROUND: Heterodimer of bone morphogenetic protein (BMP) has higher activity than BMP homodimer. Currently, there are no reports addressing comparison between BMP-4/7 heterodimer and BMP4 homodimer to induce osteogenic activity. OBJECTIVE: To compare the difference in osteogenous differentiation of bone marrow mesenchymal stem cells (BMSCs) transfected with BMP4 and BMP-4/7, and to investigate biology activity of BMP-4/7 fusion gene. METHODS: The mature peptide of BMP-4 and BMP-7 were gained by one-step RT-PCR from the human palcenta, and the BMP-4/7 fusion gene was gained through gene recombinant techniques and then transferred to pGEM plasmid. The BMP4 gene and BMP-4/7 fusion gene was cut down from the pGEM plasmid and the recombination was successfully completed in colibacillus and recombinant adeno-assosiated was produced in 293 cells. The BMSCs cell lines which could express each of the target protein w ere selected out by G418 and RT-PCR were used to certificate the expression of exogenous gene in BMSCs. Rabbit BMSCs were transfected with the recombinant adeno-assosiated virus vectors carrying BMP4 and BMP-4/7 fusion gene and cell reproductive activity detected by MTT method. The ossification of cells was evaluated by investigating the shape change of the cell ability of alkaline phosphatase and osteocalcin after transfection for 7 days. RESULTS AND CONCLUSION: We successfully constructed the recombinant adeno-assosiated virus with BMP-4 gene and BMP-4/7 fusion gene; RT-PCR results certificated the confirm expression of the exogenous gene in BMSCs. The transfection efficiency of AAV-BMP4 and AV-BMP-4/7 were 68.20% and 72.18%. MTT showed that almost all of the BMSCs which had been transfected with exogenous gene had more strong proliferative potential than the untransfected group. The cell multiplication of BMP-4 fusion gene was stronger than BMP4. There was significantly higher alkaline phosphatase and osteocalcin in AAV-BMP4 and AAV-BMP-4/7 transfection groups than untransfection group. BMP-4/7 group had a stronger osteogenous differentiation than the BMP4 group (P < 0.01). The BMP4 and AAV-BMP-4/7 fusion gene can transfect rabbit BMSCs cultured in vitro at high transfection rate, and have significant ossification of activity. The BMP-4/7 fusion gene has a stronger osteogenous differentiation than the BMP4 single gene. Related Articles | Metrics

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Experimental study on bone marrow mesenchymal stem cells inhibiting T lymphocytes proliferation in patients with aplastic anemia Liu Zeng-hui, Xiao Yang, Jiang Zu-jun, Li Li, Gao Yang, Li Yong-hua, Li Li, Wang Yao-chun, Kuang Li-ping 2011, 15 (36):  6679-6682.  doi: 10.3969/j.issn.1673-8225.2011.36.007 Abstract ( 270 )   PDF (605KB) ( 367 )   Save BACKGROUND: Previous studies have found bone marrow mesenchymal stem cells (BMSCs) have immunosuppressive effects. T lymphocytes in patients with aplastic anemia (AA) are abnormal activatory and proliferous. OBJECTIVE: To study the inhibitory effects of BMSCs on T lymphocyte proliferation in vitro in AA patients. METHODS: T lymphocytes were isolated from the peripheral blood of patients with AA and stained with carboxyfluorescein diacetate succinimidyl ester (CFDA SE), and then co-cultured with BMSCs at 1:3 ratio using closing-contact and Transwell methods. Phytohemagglutinin was added to stimulate T lymphocytes proliferation. Besides, negative and positive controls were set. RESULTS AND CONCLUSION: Flow cytometry was used to detect the T cell proliferation. CFDA SE staining analysis showed that T lymphocyte proliferation in closing-contact group was much lower than positive control (P < 0.01). T lymphocyte proliferation in Transwell group was lower than positive control (P < 0.05). Besides, T lymphocyte proliferation in closing-contact group was much lower than Transwell group (P < 0.01). BMSCs can inhibit abnormal proliferation of T cells in patients with AA possibly through closing contact and secreting some cytokines, which makes it possible to correct the immune abnormalities in AA. Furthermore, the closing-contact mechanism may play a major role in inhibiting T cell proliferation. Related Articles | Metrics

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Immunomodulatory abilities of bone marrow-derived and adipose-derived mesenchymal stem cells Zhu Xi-shan, Tai Wei-ping, Shi Wei, An Guang-yu 2011, 15 (36):  6683-6686.  doi: 10.3969/j.issn.1673-8225.2011.36.008 Abstract ( 236 )   PDF (390KB) ( 364 )   Save BACKGROUND: Do adipose-derived mesenchymal stem cells (ADMSCs) have a similar role with bone marrow mesenchymal stem cells (BMSCs) in immune regulation? OBJECTIVE: To observe the immunological characteristics of ADMSCs and BMSCs. METHODS: ADMSCs and BMSCs were isolated to detect T cell cycle, activation, inhibition and proliferation. RESULTS AND CONCLUSION: BMSCs and ADMSCs have the same function to inhibit T cell proliferation in mitogen-stimulated and mixed lymphocyte reaction of T cell proliferation in a dose-dependent manner: a strong inhibitory effect was found at a ratio of 1:2, but disappeared at a ratio of 1:100. Coculutre of BMSCs and ADMSCs could inhibit more T cells in the G0/G1 stage, and simultaneously inhibit the early activation of T cell. But the role of ADMSCs was less than that of BMSCs and ADMSCs could not inhibit T-cell apoptosis alone. Related Articles | Metrics

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Culture of rat bone marrow mesenchymal stem cells by using adherent method and shortening the time of trypsin digestion Li Shu-fa, Zhang Min 2011, 15 (36):  6687-6690.  doi: 10.3969/j.issn.1673-8225.2011.36.009 Abstract ( 278 )   PDF (648KB) ( 322 )   Save BACKGROUND: There is no uniform standard for bone marrow mesenchymal stem cells (BMSCs) isolation, purification, culture and expansion, and no specific marker for BMSCs identification. OBJECTIVE: To investigate the different isolation and culture methods of BMSCs. METHODS: Rat BMSCs were isolated and cultured by the method of frequent replacement of culture medium at early stage, controlling trypsin digestion time and plastic adherent culture. The cultured cells were induced under different conditions. The differentiated adipoccytes, osteoblasts and chondrocytes were observed. RESULTS AND CONCLUSION: Purified BMSCs can be obtained at the third generation with this isolation and culture method. The cultured cells have good proliferative potential. BMSCs can differentiate to adipocytes, osteoblasts and chondrocytes under certain conditions. Related Articles | Metrics

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Flow-cytometric method for observing the effects of pulsed electromagnetic fields on the growth of rat bone marrow mesenchymal stem cells Huang Zhao, Su Wei, Cui Xiang-rong, Qin Wan-an 2011, 15 (36):  6691-6695.  doi: 10.3969/j.issn.1673-8225.2011.36.010 Abstract ( 226 )   PDF (747KB) ( 265 )   Save BACKGROUND: Compared with the morphology, DNA electrophoresis and other methods, flow-cytometric method has more advantages on the detection of cell phenotype, proliferation rate and cell cycle of rat bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To observe and discuss the effect of specific pulsed electromagnetic fields stimulation on the proliferation and cell cycle of rat BMSCs with the flow-cytometric method (FCM). METHODS: Rat BMSCs were exposed to pulsed electromagnetic fields (frequency for 1 kHz, magnetic flux density for 0.05 mT, power density for 5 mW/cm2). BMSCs without exposure to pulsed electromagnetic fields were used as controls. Expression of CD29, CD31, CD44 CD45, CD105 were detect in the control group. The proliferation rate and cell cycle of passage 3 cells were detected at 3, 6, 9, 12 days. RESULTS AND CONCLUSION: CD29, CD44, CD105 in the 3rd passage non-stimulated BMSCs was positively expressed and the CD31, CD45 was negatively expressed (P < 0.05). The survival rate of passage 3 BMSCs and percentage of S phase following intervention of pulsed electromagnetic fields were greater than those in the control group (P < 0.05). These results indicated that the specific pulsed electromagnetic fields can promote the growth and proliferation of BMSCs to some extent. Related Articles | Metrics

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pcDNA3.1-osteogenic growth polypeptide eukaryotic expression vector in bone marrow mesenchymal stem cells An Gang, Lü Song-cen, Guo Ya-shan, Xue Zhen, Deng Qiu-kui 2011, 15 (36):  6696-6700.  doi: 10.3969/j.issn.1673-8225.2011.36.011 Abstract ( 344 )   PDF (492KB) ( 284 )   Save BACKGROUND: Osteogenic growth polypeptide (OGP) had clear effect on promoting osteoblast proliferation, differentiation and mature. OBJECTIVE: To explore the expression of OGP gene, which was transfected into rabbit bone marrow mesenchymal stem cells (BMSCs) and to evaluate the effects of OGP on differentiation of rabbit BMSCs. METHODS: pcDNA3.1-OGP was constructed using gene cloning and recombination techniques. Rabbit BMSCs were transfected with pcDNA3.1-OGP mediated by lipofectamine 2000. The transfection positive cell clones were selected with G418. The expression of OGP gene was detected using reverse transcription-polymerase chain reaction analysis on an mRNA level. Differentiation of pcDNA3.1-OGP transfected BMSCs into osteoblast lineage was observed. RESULTS AND CONCLUSION: The pcDNA3.1-OGP plasmid was constructed successful and OGP expression was detected in rabbit BMSCs. Hydroxyproline content was increased, and alkaline phosphatase activity was also increased. These indicate that pcDNA3.1-OGP transfected BMSCs expressed OGP, and could differentiate into osteoblast lineage. Related Articles | Metrics

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Umbilical cord mesenchymal stem cells effect on inflammatory responses of rats with type Ⅱ collagen-induced arthritis Li Hui, Gu Jian, Lin Chuan-ming, Ma Li, Shen Lian-jun, Wu Wei, Wang Zhong-qiang 2011, 15 (36):  6701-6704.  doi: 10.3969/j.issn.1673-8225.2011.36.012 Abstract ( 285 )   PDF (574KB) ( 333 )   Save BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) can express various kinds of embryonic stem cell-specific molecular markers, and be characterized as good differentiation potential, strong proliferative capacity, and low immunogenicity. OBJECTIVE: To observe the interventional effect of UC-MSCs on immune inflammatory pathological process of rheumatoid arthritis.  METHODS: CIA rat models were established and divided into three groups. At 4 weeks after modeling, UC-MSCs were injected via the tail vein in the UC-MSCs group, and normal saline was injected in the model group. RESULTS AND CONCLUSION: The levels of interleukin-1 (IL-1), IL-6, IL-18, tumor necrosis factor alpha, vascular cell adhesion molecule and neutrophil cell surface expression of CD11b were higher in the model group than the normal group (P < 0.05) at 7 and 35 days after treatment. Those factor levels in the UC-MSCs group were significantly lower than those in the model group after treatment (P < 0.05), those at 7 days were higher than at 35 days (P  < 0.05). UC-MSCs can obviously inhibit the release of inflammatory cytokines and abnormal activation of endothelial cells, and relieve immune and inflammatory reactions of rheumatiod arthritis. Related Articles | Metrics

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5-azacytidine induces the differentiation of human umbilical cord derived mesenchymal stem cells into cardiomyocytes Ruan Zhong-bao, Yang Xiang-jun, Chen Ge-cai, Zhu Li, Li Wei, Yang Bing, Ouyang Xi, Pan Shao-hui 2011, 15 (36):  6705-6708.  doi: 10.3969/j.issn.1673-8225.2011.36.013 Abstract ( 192 )   PDF (1312KB) ( 302 )   Save BACKGROUND: 5-azacytidine (5-aza) can induce the differentiation of human umbilical cord derived mesenchymal stem cells (hUCMSCs) into cardiomyocytes. OBJECTIVE: To investigate the cellular mechanism underlying the differentiation of hUCMSCs into cardiomyocytes induces by 5-aza. METHODS: hUCMSCs were isolated and purified by adherent method. hUCMSCs at passage 3 were induced to differentiate into cardiomyocytes by 5-aza. RESULTS AND CONCLUSION: Spindle-shaped hUCMSCs were presented from human umbilical cord tissue by adherent culture method. hUCMSCs treated with 5-aza had long cytoplasmic process 1-2 weeks after induction, and they were connected with adjacent cells forming myotube-like structures 3-4 weeks after induction. hUCMSCs treated with 5-aza were stained positively for cTnI 4 weeks after induction. Statistically higher level expression of Nkx2.5 and GATA 4 mRNA in induced group was observed in comparison with those of control group (P < 0.05). These indicated that hUCMSCs can be induced to differentiate into cardiomyocytes by 5-aza in vitro through up-regulating the expression of Nkx2.5 and GATA4. Related Articles | Metrics

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Differentiation of human amnion derived-mesenchymal stem cells into hepatocytes in rat injured liver  Gong Li-ming, Fang Ning, Chen Dai-xiong, Wan Wei-hong, Zhang Tao, Liu Zu-lin, Yu Li-mei, Zhao Chun-hua 2011, 15 (36):  6709-6713.  doi: 10.3969/j.issn.1673-8225.2011.36.014 Abstract ( 206 )   PDF (1396KB) ( 289 )   Save BACKGROUND: Human amnion derived-mesenchymal stem cells (hAD-MSCs) can differentiate into hepatocyte-like cells in certain conditions in vitro. It needs to further investigate whether they can differentiate into hepatocytes in damaged liver. OBJECTIVE: To study the survival and differentiation of hAD-MSCs in vivo in rat injured liver. METHODS: hAD-MSCs were isolated from human amnion treated with trypsin and collagenase. Forty healthy and clean grade female SD rats underwent intraperitoneal injection of D-galactosamine diluted in normal saline at the dose of 400 mg/kg body weight, to establish the model of liver injury, and then divided into hAD-MSCs group and control group stochastically. Twenty-four hours after modeling, 50 μL cell suspension (approximately 1×106 cells suspended in L-DMEM) of hAD-MSCs were injected slowly into the left, middle and right hepatic lobe respectively with micro-syringe, while equivalent volume of L-DMEM injected into the control group. RESULTS AND CONCLUSION: ①Freshly isolated hAD-MSCs expressed CD29, CD44 and CD166; immunofluorescence staining showed that vimentin was positive in hAD-MSCs, while cytokeratin 19 was negative. ②There was no significant difference after transplanting hAD-MSCs into injured liver between the hAD-MSCs group and the control group, remaining acute hepatic necrosis. ③hAD-MSCs implanted into injured liver were located in hepatic lobules at day 7 after transplantation, which expressed CK19; and the cells expressed CK18 at day 14, and also human Alb at day 21. hAD-MSCs xenografted to rat injured liver can survive and differentiate into hepatocytes, suggesting that hAD-MSCs transplantation may have potential applications for treating clinical liver injury diseases. Related Articles | Metrics

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Observation of morphology and distribution of CD44+/CD166+ colorectal cancer stem cells by tissue microarray Luo Yan, Liu Bin, An Ning, Dong Liang, Qian Zhen, Zhang Yong-dong 2011, 15 (36):  6714-6717.  doi: 10.3969/j.issn.1673-8225.2011.36.015 Abstract ( 373 )   PDF (1075KB) ( 317 )   Save BACKGROUND: More and more research employs CD44 as a specific marker of colorectal cancer stem cells. OBJECTIVE: To investigate the quantity, location and distribution of colorectal cancer stem cells (Co-CSCs) CD44+/CD166+ in primary colorectal carcinoma. METHODS: A totally of 61 cases of human colorectal carcinoma and 10 cases of normal mucosa, 18 cases of adenoma were collected and made into three tissue microarrays each containing of 42 dots respectively. RESULTS AND CONCLUSION: The results of double-label immunohistochemical staining demonstrated there was no CD44+/CD166+ cell in normal intestine mucosa, a very small amount of CD44+/CD166+ cells in adenoma, also double-positive cells could be seen in colorectal carcinoma, the number of double-positive cells was rare and the cells were scattered or distributed focally along the basement of gland basal side. The cells with scarcely cytoplasm were square, and its nucleus was oval or high cylindrical, deep stained and homogeneous. The quantity of double-positive cells was negatively correlated with the differentiation of colorectal carcinoma. The more depth was infiltrated, the more quantity of double-positive cells was observed. To observe the number, location, distribution and morphology of Co-CSCs can contribute to the diagnosis and prognostic evaluation of colorectal carcinoma. Related Articles | Metrics

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Isolation and aipogenic differentiation of gastric cancer stem cells Kuang Tao, Wang Lei, Song Wen, Meng Dong-mei 2011, 15 (36):  6718-6721.  doi: 10.3969/j.issn.1673-8225.2011.36.016 Abstract ( 290 )   PDF (1221KB) ( 283 )   Save BACKGROUND: Whether cancer stem cells are present in all tumors is still controversial. OBJECTIVE: To isolate and identify cancer stem cells from human gastric cancer cells, and to explore the mechanism of gastric cancer. METHODS: Gastric cancer cells were primarily cultured and CD44 and CD29 positive expression rates were detected by flow cytometry to sort positive cells for adipogenic induction. RESULTS AND CONCLUSION: CD44 and CD29 positive percentages in gastric cancer cells were 5.67% and 5.53%, respectively, indicating that CD44 and CD29 positive cells had an adipogenic capacity. It is proved that there are tumor stem cells with differentiation capacity in gastric cancer cells. Related Articles | Metrics

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Effect of transforming growth factor beta 1 on differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells Hu He, Han Cheng-long, Jiang Chao, Shen Hong-tao, Liu Yang, Yan Feng, Yu Chang-shui, Yong Mei 2011, 15 (36):  6722-6726.  doi: 10.3969/j.issn.1673-8225.2011.36.017 Abstract ( 189 )   PDF (1420KB) ( 275 )   Save BACKGROUND: Transforming growth factor beta 1 (TGF-β1) is a first selected growth factor in differentiation of bone marrow mesenchymal stem cells (BMSCs) into nucleus pulposus (NP)-like cells. Suitable concentration can induce proliferation and differentiation of BMSCs. OBJECTIVE: To observe the effects of different-concentration TGF-β1 on differentiation of BMSCs into NP-like cells, and to optimize culture conditions. METHODS: BMSCs were separated and purified from adult rat femur bone marrow in vitro. The third-generated BMSCs were taken and induced with different concentrations of TGF-β1-contained in HG-DMEM culture medium in the experimental group, while in control group HG-DMEM medium containing 10% fetal bovine serum was used in general state. Immunohistochemical method was used to detect collagen type Ⅱ, and RT-PCR to detect the mRNA expression of collagen type Ⅱ and proteoglycan. RESULTS AND CONCLUSION: The proteoglycans and collagen type Ⅱ of BMSCs was significantly higher than other experimental and control groups when TGF-β1 concentration was 10 μg/L (P < 0.01). The expression of proteoglycans at 14 days were higher than that at 3, 7, 21 days in the experimental groups (P < 0.01). The expression of proteoglycans and collagen type Ⅱ in the control group was negative. TGF-β1 can increase the number of differentiated BMSCs into NP-like cells at concentration of 10 μg/L, and make BMSCs play a greater treatment efficiency. Related Articles | Metrics

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In vitro combination of different-concentration hepatocyte growth factor and epidermal growth factor induces differentiation of rabbit bone marrow mesenchymal stem cells into hepatocytes Luo Wei, Xiao En-hua, Luo Jian-guang, Shang Quan-liang, Wu Yu-zhi, Li Yan-hui, Tan Yan 2011, 15 (36):  6727-6731.  doi: 10.3969/j.issn.1673-8225.2011.36.018 Abstract ( 208 )   PDF (1295KB) ( 312 )   Save BACKGROUND: Studies have confirmed that in a variety of micro-environments, bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into mature hepatocytes, but the inducible condition and differentiating rate are inconclusive. To select the appropriate inducer and its concentration is particularly important. OBJECTIVE: Through study on the differentiation of rabbit BMSCs into hepatocytes in vitro induced by different concentrations of hepatocyte growth factor (HGF) and epidermal cell growth factor (EGF), to find the best concentration combination of HGF and EGF. METHODS: BMSCs were induced with different concentrations of HGF and HGF. The morphological changes of cells were observed. In different culture periods, the hepatic surface phenotype including alpha fetoprotein (AFP), albumin (ALB) and synthesis function of hepatocytes were determined. RESULTS AND CONCLUSION: After prolonged culture, hepatocyte-like cells were seen. Until day 7, AFP was positive, but reduced in the following days (P < 0.05), and after that, there were no difference among all groups (P > 0.05). Until day 14, ALB expression was positive, and increased with the culture time (P < 0.05). The higher the concentration of cell growth factors, the higher the number of positive cells (P< 0.05). The concentration of ALB in the supernatant in the early stage (9-15 days) showed a concentration-dependent manner, and reached the peak at 18 or 21 days. Then the concentration began to decreased, and there were no difference (P > 0.05). The high concentrations of HGF and EGF may enhance the rate of differentiation, and the best combination is HGF 60 ug/L and EGF 4.5 mg/L. Related Articles | Metrics

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Effects of simvastatin on Wnt and bone morphogenetic protein 2 signaling pathway during osteoblast differentiation of bone marrow stromal cells Jin Yun-qiao, Zhang Liu, Tian Fa-ming, Zhang Lei, Cheng Tan, Liu Xiao-ning 2011, 15 (36):  6732-6736.  doi: 10.3969/j.issn.1673-8225.2011.36.019 Abstract ( 289 )   PDF (1545KB) ( 305 )   Save BACKGROUND: Simvastatin can promote osteoblast differentiation of human or rat bone marrow stromal stem cells cultured in vitro. But the mechanism remains unclear. OBJECTIVE: To investigate the effects of simvastain on expression of factors related to Wnt and bone morphogenetic protein 2 (BMP-2) signaling pathway during osteoblast differentiation of bone marrow stromal cells. METHODS: Bilateral femur and tibia bone marrow was harvested from 6-week-old female Sprague-Dawley rats for in vitro culture of osteoblasts. In the SIM group, 10-7 mol/L simvastain was added, and in the control group, the same amount of dehydrated alcohol and PBS was added. After 14 days of culture, alkaline phosphatase staining was performed. At 28 days, von Kossa staining was applied to observe osteoblastic differentiation and mineralization. At 14 and 21 days, immunofluorescent cytochemical staining was used to detect β-catenin, Smad1/5, Cbfa1 expression and distribution in osteoblasts.  RESULTS AND CONCLUSION: Cells in each group were differentiated into osteoblast lineage, as shown by alkaline phosphatase activity and extracellular matrix mineralization. Simvastatin can significantly upregulate alkaline phosphatase expression during the osteoblast differentiation of bone marrow stromal cells. The expression of β-catenin, Smad1/5, and Cbfa1 was significantly increased in the SIM group than in the control group. The above-mentioned expression in the SIM group accumulated in the nuclear more than in the cytoplasm. These results showed that the mechanism by which simvastatin promotes osteoblast differentiation of bone marrow stromal stem cells may be partially related to expression and distribution of factors regulating Wnt and BMP-2 signaling pathway. Related Articles | Metrics

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Stem cell factor and granulocyte colony-stimulating factor affect the migration of bone marrow mesenchymal stem cells in rats with myocardial infarction Wu Zu-chang, Wu Guo-cai, Wu Zhi-ming, Xiong Dan, Lü Jun-ting, Yang Zhi-gang 2011, 15 (36):  6737-6740.  doi: 10.3969/j.issn.1673-8225.2011.36.020 Abstract ( 246 )   PDF (1062KB) ( 348 )   Save BACKGROUND: Only 1%-5% of the transplanted mesenchymal stem cells (MSCs) home to the myocardial infarction area through peripheral venous transplantation. OBJECTIVE: To investigate the effects of stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) on MSCs homing. METHODS: MSCs were isolated from SD rats, and purified by adhesive-screening method. MSCs at passages 3-5 were assigned to 4 groups: groups of MSCs, SCF, G-CSF and SCF+G-CSF. RESULTS AND CONCLUSION: Under fluorescence microscope, the number of MSCs migrating to the myocardial infarction region had significant difference among MSCs group, SCF group and G-CSF group (P > 0.05), but SCF+G-CSF group had the higher number of MSCs migrating to the myocardial infarction region than other three groups (P < 0.05). Immunofluorescence histochemistry showed that some MSCs express cardiac-specific protein cTnI. Combination of SCF and G-CSF can promote MSCs homing, and induced by microenvironment in vivo, MSCs can convert into cardiomyocyte-like cells. Related Articles | Metrics

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Effect of bone marrow stromal cells in combination with exogenous cytokines on antigen expression of cord blood mononuclear cells Zeng Jin-long, Zeng Lian-kun, Wang Zhi-chao, Zhang Hai-liang, Mao Ping 2011, 15 (36):  6741-6744.  doi: 10.3969/j.issn.1673-8225.2011.36.021 Abstract ( 247 )   PDF (1095KB) ( 220 )   Save BACKGROUND: In vitro hematopoietic culture system of fetal bone marrow stromal cells (BMSCs) combined with exogenous cytokines has been established. It remains to verify whether this culture system can improve the proliferation of hematopoietic cells at various developmental stages. OBJECTIVE: To investigate the effects of the culture system containing BMSCs in combination with exogenous cytokines on umbilical cord blood (UCB) mononuclear cells (MNCs) expressing CD133 and CD34. METHODS: MNCs separated from UCB were cultured in a serum-free system with BMSCs or exogenous cytokines or both of them. On days 0, 6, 10 and 14, total nucleated cells (TNCs) were counted, CD133+, CD34+, and CD133+CD34+cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay.  RESULTS AND CONCLUSION: The number of TNCs was remarkably more in BMSCs and cytokine group compared with the other two groups. The expression of CD133+ and CD133+CD34+ cells were higher in BMSCs and cytokine group than the other two groups at days 6 and 10. The ratio of CD133+, CD34+ and CD133+CD34+ cells was significantly more in the system containing BMSCs. BMSCs in combination with cytokines can expand UCB-MNCs and CD133+, CD34+, CD133+CD34+ cells effectively. BMSCs exert an important role in remaining primitive characteristics of hematooietic stem/progenitor stem cells. Related Articles | Metrics

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Effects of cardiotrophin-1 on glutamate induced apoptosis of neural stem cells Shen Dong-lin, Shu Xiao-mei, Li Zhen-hong, Chen Xue-mei, Du Shu-zhen 2011, 15 (36):  6745-6748.  doi: 10.3969/j.issn.1673-8225.2011.36.022 Abstract ( 270 )   PDF (1188KB) ( 274 )   Save BACKGROUND: Cardiotrophin-1 (CT-1) appertains to interleukin-6 cytokine family, which is found in recent years to have an important protective role in the central nervous system. OBJECTIVE: To observe the protective effects of CT-1 on the apoptosis of neural stem cells (NSCs) induced by glutamate. METHODS: Hippocampal neural stem cells isolated from newborn Wistar rats were passaged and divided into 3 groups: glutamate, CT-1 (transfected with Adv CT-1) and control groups. NSCs survivals were estimated by MTT colorimetric method, and the apoptosis of NSCs was detected by means of TdT-Mediated dUTP nick end labeling (TUNEL). RESULTS AND CONCLUSION: Glutamate treatment alone induced cell death that was associated with obvious morphological changes, while these changes were inhibited by transfection of CT-1 into NSCs. At 12, 24, 48 and 72 hours after treatment with glutamate, the absorbance values of MTT in the lutamate group were significantly lower than those in the CT-1 group, the apoptosis rates of NSCs in the glutamate group were significantly higher than those in the CT-1 group. At the same time, the absorbance values of MTT in the control group were significantly higher than those in the glutamate group and CT-1 group, the apoptosis rates of NSCs were significant lower than those in the glutamate group and CT-1 group. These indicated that CT-1 has a protective effect on NSCs injured by glutamate, can inhibit NSCs apoptosis and improve NSCs survival. Related Articles | Metrics

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Endothelial progenitor cells pre-cultured with stromal cell derived factor 1 alpha transplantation to treat hind limb ischemia in diabetic rats Wang Ning, Wang Hu-guo, Jiang Tao, Duan Ming-jun, Xie Zi-jing 2011, 15 (36):  6749-6752.  doi: 10.3969/j.issn.1673-8225.2011.36.023 Abstract ( 310 )   PDF (1428KB) ( 280 )   Save BACKGROUND: Stromal cell derived factor 1 alpha (SDF-1α) has been proved to promote endothelial progenitor cells (EPCs) homing. OBJECTIVE: To explore the effects of SDF-1α pre-cultured EPCs from autologous bone marrow on hind limb ischemia in diabetic rat models. METHODS: Thirty male Wistar rats (successful model preparation in 25 rats) were randomly divided into 3 groups: normal saline control group (A group), EPCs transplantation in alloxan-induced diabetes rats (B group), EPCs cultured by SDF-1α transplantation in alloxan-induced diabetes rat (C group). RESULTS AND CONCLUSION: MTT results showed SDF-1α could remarkably improve the proliferative ability of EPCs (P < 0.01). Angiography showed that angiogenesis index was significantly higher in the C group at the 28th day (P < 0.05). EPCs pre-cultured with SDF-1α have a promoted effect to reform the hind limb ischemia condition in diabetic rats; blood-supply mainly comes from new vessels and /or virgin minute blood vessel’s compensation thickening. Related Articles | Metrics

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Improvements of ventricular fibrillation threshold and cardiac electrophysiological stability in myocardial infarction following cardiac stem cells transplantation Wang Tong, Zheng Shao-xin, Zhou Chang-qing, Weng Yin-lun, Wen Zhu-zhi, Huang Hui, Wang Jing-feng 2011, 15 (36):  6753-6756.  doi: 10.3969/j.issn.1673-8225.2011.36.024 Abstract ( 272 )   PDF (543KB) ( 326 )   Save BACKGROUND: Whether transplanted cardiac stem cells (CSCs) can improve ventricular fibrillation threshold (VFT) and whether severe malignant ventricular arrhythmia is induced in the myocardial infarction model are still unclear. OBJECTIVE: To clarity the electrophysiological characteristics and VFT in rats with myocardial infarction by treatment with allogenic CSCs. METHODS: Left anterior descending coronary arteries of SD rats were ligated to induce myocardial infarction. Two weeks later, animals were randomized to receive 5×106 CSCs labeled with PKH26 in phosphate buffer solution (PBS) or PBS alone as a placebo by injection into the ischemic zone in the anterior ventricular free wall. Six weeks after the CSCs or PBS injection, electrophysiological characteristics and VFT were measured at the infarct area. Labeled CSCs were observed in 5 µm cryostat sections from each harvested heart. RESULTS AND CONCLUSION: Compared with the PBS group, the unipolar electrograms activation recovery time dispersions were shorter in the CSCs group, malignant ventricular arrhythmias were significantly less inducible in the CSCs group, the VFTs were greatly improved in the CSCs group. Labeled CSCs were identified in the infarct marginal zone and expressed Connexin-43 and α-sarcomeric actin. CSCs can modulate the electrophysiological abnormality and improve the VFT in rats with myocardial infarction treated with allogenic CSCs and CSCs express muscle and connexin markers in vivo. Related Articles | Metrics

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Adoptive immunotherapy for leukemia in mice after autologous bone marrow mixed with H-2 haploidentical allogeneic bone marrow transplantation Wang Cun-bang, Bai Hai, Xi Rui, Zhang Qian, Zhou Jin-mao, Zhao Qiang, Pan Yao-zhu 2011, 15 (36):  6757-6761.  doi: 10.3969/j.issn.1673-8225.2011.36.025 Abstract ( 278 )   PDF (640KB) ( 263 )   Save BACKGROUND: Auto hemopoietic stem cell transplantation (Auto-HSCT) has a high relapse rate in acute leukemia; allo-HSCT has a high incidence of transplant-related mortality. It may increase curative effect when leukemia patients are administered adoptive immunotherapy post mixed-HSCT. OBJECTIVE: To explore the curative effect of using donor lymphocyte infusion combined with interleukin-2 (DLI+IL-2) after autologous bone marrow mixed with H-2 haploidentical allogeneic bone marrow transplantation (MBMT) in mice with leukemia. METHODS: Leukemia models were prepared with Balb/c mice which were irradiated 3 Gy by linear accelerator and injected K562 (GFP+/NeoR+) or K562 (GFP-/NeoR-) cells 5×105 into caudal vein and divided into leukemia model group, irradiated leukemia model group, MBMT group, and autologous bone marrow transplantation (ABMT) group. 6 Gy irradiation was performed after 7 days; the mice were treated with ABMT or MBMT respectively. Mice of MBMT group mixed with 1/10 of H-2 haploidentical allogeneic bone marrow cells underwent IL-2 or combination of DLI treatment. Peripheral blood and bone marrow cell morphous of mice were examined; cell subsets, GFP and NeoR gene in peripheral blood, and liver, spleen homogenate cells, and NeoR gene were detected after 4 weeks. RESULTS AND CONCLUSION: All of mice in leukemia model group died of bone marrow hematopoietic failure within 20 days; mice in irradiated leukemia model group died of hematopoietic failure within 14 days. Varying amounts of non-leukemic of mice survived for more than 28 days between ABMT group and MBMT group. UsingIL-2 treatment after MBMT and ABMT can promote long term disease free survival of mice with leukemia, and which combined with DLI can further improve long term disease free survival of mice with leukemia. Related Articles | Metrics

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Effects of complement 5a receptor antagonists combined with Schwann cells transplantation on recovery of electrophysiological and neurological function in rats with spinal cord injury Wang Yu-cheng, Zhou Jiang-bo 2011, 15 (36):  6762-6766.  doi: 10.3969/j.issn.1673-8225.2011.36.026 Abstract ( 324 )   PDF (602KB) ( 253 )   Save BACKGROUND: Complement 5a (C5a) by enhancing the early inflammatory response after spinal cord injury participates in the spinal cord secondary injury. C5a receptor antagonists can effectively block this process. OBJECTIVE: To investigate the effect of Schwann cells transplantation and C5a receptor antagonists on repair of rat spinal cord injury. METHODS: Eighty healthy female Wistar rats were randomly divided into 4 groups: control group, cell transplantation group, C5a receptor antagonist group and combination group, 20 rats in each group. RESULTS AND CONCLUSION: The outcome of combination group was superior to the cell transplantation group and the C5a receptor antagonist group, and the cell transplantation group and the C5a receptor antagonist group was superior to the control group (P < 0.05). Cell transplantation group compared with the C5a receptor antagonist was no significant difference (P > 0.05). The SRY expression in cell transplantation group and combined group, in contrast, no SRY gene were detected in control group and the C5a receptor antagonist group. The number of HRP-positive nerve fibers: the combination group > cell transplantation group and C5a receptor antagonist group > control group and the difference between the groups was significant (P < 0.01). Somatosensory evoked potential, motor evoked potential latency, and amplitude were significantly better in the combination group than the other three groups (P < 0.05 or P < 0.01). Schwann cell transplantation combined with the C5a receptor antagonist can promote the regeneration of nerve synapses in rats with spinal cord injury, and to improve their motor function, and electrophysiological function. Related Articles | Metrics

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Effects of Bing’s giant salamander containing serum on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells Xie Xing-wen, Xu Wei, Li Ning 2011, 15 (36):  6767-6771.  doi: 10.3969/j.issn.1673-8225.2011.36.027 Abstract ( 267 )   PDF (724KB) ( 349 )   Save BACKGROUND: Bing’s giant salamander can promote fracture healing, but the mechanism and active ingredients are rarely reported. OBJECTIVE: To investigate the effect of Bing’s giant salamander containing serum on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). METHODS: Rats were given Bing’s giant salamander by high, moderate and low doses and their serum were obtained after 7 days. The control serum was obtained by giving equal volume physiological saline. Rat BMSCs were obtained from Wistar rats and screened by the adhesive method. The serum was supplemented into the culture medium of rat BMSCs by different concentration. The proliferation of rat BMSCs was analyzed by MTT assay. The osteogenic differentiation markers including Osterix expression, alkaline phosphatase (ALP) activity, calcium deposition and mineralized bone modulus were compared among the exposed groups and the control at different time point. RESULTS AND CONCLUSION: Moderate Bing’s giant salamander containing serum showed a stronger stimulation of cell proliferation and significantly enhanced the osteogenic differentiation of rat MSCs, indicated by significantly improving ALP activity, calcium deposition, OSX expression and the number of mineralized bone nodules compared to the control and other groups (P < 0.05). Related Articles | Metrics

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Effects of Zhichan Decoction on intracerebral dopamine and metabolites after neural stem cells transplanted in Parkinson disease rats Shi Hui-fen, Lu Yu, Song Jie, Li Wen-tao 2011, 15 (36):  6772-6775.  doi: 10.3969/j.issn.1673-8225.2011.36.028 Abstract ( 252 )   PDF (537KB) ( 318 )   Save BACKGROUND: How to make the transplanted neural stem cells differentiated into more dopaminergic neurons, so as to accomplish the plerosis of injured dopaminergic neuron. This problem solving will be the breakthrough progress for the treatment of Parkinson disease (PD). OBJECTIVE: To study the effects of Zhichan Decoction on substantia nigra dopamine (DA) and metabolites, and the survival and directional differentiation after neural stem cells were transplanted from metabolic pathway in PD rats. METHODS: PD rats models were established by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine. The contents of DA, homovanillic acid (HVA) and 3, 4-dihydroxy phenyl acetic acid (DOPAC) were measured with high-performance liquid chromatography (HPLC).  RESULTS AND CONCLUSION: Zhichan Decoction could increase the content of DA and DOPAC. But there were no effects on HVA. The Zhichan Decoction has the effect to promote the neural stem cells differentiating into dopaminergic neurons in PD rats; At the same time, it could inhibit DA breaks down, so that the therapeutic effect was achieved. Related Articles | Metrics

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Differentiation of murine embryonic stem cells into hematopoietic stem cells induced by stromal cells in human aorta-gonad-mesonephros region and hematopoietic reconstitution in vivo Cai Yun, Zhang Xu-chao, Chen Hui-qin, Huang Shao-liang 2011, 15 (36):  6776-6780.  doi: 10.3969/j.issn.1673-8225.2011.36.029 Abstract ( 276 )   PDF (743KB) ( 319 )   Save BACKGROUND: Up to now it has been confirmed that under proper conditions of hematopoietic growth factors, feeder layers of stromal cells from hematopoietic niches and their conditioned media, embryonic stem cells (ESCs) can be induced into hematopoietic stem cells (HSCs). OBJECTIVE: To direct murine ESCs into HSCs in vitro by the support of human aorta-gonad-mesonephros (AGM) region stromal cells, and to compare the function of hematopoietic reconstitution in vivo by different routes of HSCs transplantation in mice. METHODS: Firstly, E14 murine ESCs were induced into embryoid body (EB). Then the cells from EB were further co-cultured with human AGM region stromal cells in Transwell non-contact system for 6 days. The induced EB cells were collected for Sca-1+c-Kit+ cells analysis by flow cytometry, checked for teratoma formation and transplanted to BALB/C female mice conditioned with lethal dose 60Co γ-ray irradiation. The recipient mice were divided into four groups at random, including intravenous injection group, bilateral tibia intra-bone marrow (IBM) injection group, irradiation control group and normal control group. The survival rates, reconstitution of hematopoietic and engraftment of donor cells of different groups were monitored. RESULTS AND CONCLUSION: Sca-1+c-Kit+ cells in EB cells after co-cultured with human AGM region stromal cells accounted for (13.12±1.30)%. Teratoma could be detected in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region stromal cells, while there was no teratoma in the mice after IBM injection of that cells. The recipients in intravenous injection group all died. The survival rate was 55.6% in IBM group, in which the peripheral blood cell count was near to the normal at day 21 after transplantation and Sry gene copies from donor could be detected. It suggested that feeder cells from human AGM region can induce directed differentiation of murine ESCs into HSCs which can reconstruct hematopoiesis in vivo without teratoma by IBM administration. Related Articles | Metrics

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Harvesting peripheral blood stem cells from major ABO mismatched donors using COM.TEC blood cell separator Wang He-hua, Chen Hui-zhen, Zou Wai-yi, Xu Duo-rong, Chen Yun-xian 2011, 15 (36):  6781-6784.  doi: 10.3969/j.issn.1673-8225.2011.36.030 Abstract ( 309 )   PDF (592KB) ( 273 )   Save BACKGROUND: Approximately 1/3 allogeneic peripheral blood stem cells (PBSCs) transplantation treatments are performed between ABO mismatched donors-recipients pairs. OBJECTIVE: To evaluate the feasibility of PBSCs collection in major ABO mismatched donors with COM. TEC blood cell separator and infusion to the recipients without removal of erythrocytes from the PBSCs graft. METHODS: Twenty-seven HLA-identical sibling donors received priming with granulocyte colony stimulating factor (G-CSF)    5 μg/kg/d via subcutaneous injection for 5 days. PBSCs collection started on day 5 using COM.TEC blood cell separator. The procedure processed 3.0-3.5 times of the total blood volume (TBV). For major ABO mismatched donors, specialized harvesting strategies of adjusting the volume of buffy coat and maintaining adequate and stable flow rate during apheresis were to diminish red blood cells (RBCs) contents of PBSCs. RESULTS AND CONCLUSION: Of the 27 donors, there were 11 major ABO-incompatible and 16 ABO-identical. The volumes of PBSCs graft and RBCs harvested in major ABO-incompatible group were significantly lower than those of ABO-identical group (P　< 0.05). The number of mononuclear cells and CD34+ cells per recipient body weight did not statistically differ between the two groups (P > 0.05). All recipients reconstituted hematopoiesis following allo-PBSCT. Mean time to neutrophil and platelet engraftment was no significant difference between ABO-identical and major incompatible transplantaions (P > 0.05). PBSC harvesting from major ABO mismatched donors can be successfully performed with COM.TEC blood cell separator. Adjustment the volume of buffy coat and maintenance of adequate and stable flow rate during procedure can reduce RBC contamination for prevention of immune hemolysis; meanwhile collecting a sufficient number of CD34+ cells to ensure prompt engraftment post-transplantation. Related Articles | Metrics

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In vitro differentiation of human amniotic mesenchymal stem cells into vascular endothelial cells Shi Jing-jing, Zhu Dong-xiao, Jin Wei-dong, Wang Xiao-yan, Zhuang Rui-juan, Li Bai-hong, Miao Zong-ning 2011, 15 (36):  6785-6788.  doi: 10.3969/j.issn.1673-8225.2011.36.031 Abstract ( 250 )   PDF (1119KB) ( 230 )   Save BACKGROUND: Whether human amniotic mesenchymal stem cells can differentiate into corresponding target cells after transplantation into different types of tissues? OBJECTIVE: To investigate the feasibility of differentiation from human amniotic mesenchymal stem cells into vascular endothelial cells in vitro induced by vascular endothelial cell growth factor. METHODS: Human amniotic mesenchymal stem cells were isolated, and then characterized by flow cytometry using a panel of monoclonal antibodies. Differentiation into endothelial-like cells was induced by cultivation of confluent cells in the presence of 2% fetal calf serum and 50 μg/L vascular endothelial growth factor. We characterized these cells by immunofluorescence staining for endothelial-specific markers KDR and the von Will brand factor. RESULTS AND CONCLUSION: Isolated human amniotic mesenchymal stem cells 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. Immunofluorescence staining analysis of the confluent cells in situ showed positive for endothelial-specific markers like KDR and the von Will brand factor. The human amniotic mesenchymal stem cells have the capability to differentiate into vascular endothelial cells in vitro. Related Articles | Metrics

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Isolation, culture and identification of rat embryonic mesencephalic dopaminergic neurons Wei Xiao-bing, Deng Xing-li, Wang Ying-li, Yang Zhi-yong, Li Zhi-gao, Liu Ru-en, Li Yu 2011, 15 (36):  6789-6792.  doi: 10.3969/j.issn.1673-8225.2011.36.032 Abstract ( 318 )   PDF (1155KB) ( 273 )   Save BACKGROUND: Nerve cell transplantation therapy is one of the most promising methods to cure Parkinson’s disease. OBJECTIVE: To investigate the culture, isolation and identification of rat embryo mesencephalic dopaminergic neurons. METHODS: The mesencephalon was dissected from rat embryo at embryonic day 14 (E14). The brain tissue was triturated into a fine single-cell suspension. The cells were cultured with the medium containing Neurobasal, B27 supplement, β-mercaptoethanol and fetal bovine serum. The cells were identified with RT-PCR as well as immunocytochemistry, respectively. RESULTS AND CONCLUSION: The cells survived well in the medium. RT-PCR and immunocytochemistry results showed mostly all neurons expressed molecular markers for dopaminergic neurons. The dopaminergic neurons derived from rat embryonic mesencephalon were successfully isolated and cultured, which will provide the foundation for the further research about cell therapy of Parkinson’s disease. Related Articles | Metrics

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Effect of frozen pretreatment temperature on bioactivity of embryonic rabbit’s Schwann cells Dai Xiu-song, Wu Ming, Zhang Chang-chun, Xu Wei, Xiao Yu-zhou 2011, 15 (36):  6793-6796.  doi: 10.3969/j.issn.1673-8225.2011.36.033 Abstract ( 274 )   PDF (1306KB) ( 249 )   Save BACKGROUND: Schwann cells are considered as an important bridge material for peripheral nerve repair, and the activity of Schwann cells is a key to whether the repair of peripheral nerve injury succeeds or not. OBJECTIVE: To investigate the effect of different frozen pretreatment temperatures on the bioactivity of embryonic rabbit’s Schwann cells preserved at ultra-low temperature. METHODS: The embryonic rabbit’s Schwann cells were frozen at -30 ℃, -35 ℃, -40 ℃, -45 ℃, -50 ℃, -55 ℃, -60 ℃, -65 ℃ as well as -70℃ respectively, treated by a cryoprotectant, 10% dimethyl sulfoxide, at the same time, preserved in liquid nitrogen for 48 hours, and cultured for 48 hours after rapid rewarming. Morphological changes of Schwann cells were observed under transmission electron microscopy (TEM). Bioactivity of Schwann cells was assessed by MTT and 3H-TdR incorporation test assays. RESULTS AND CONCLUSION: The ultrastructure of Schwann cells in the group of -45 ℃ remained intact under TEM; however, other experimental cells exhibited the different appearances of cryodamage such as swelling organelles, necrosis in the cytoplasm. Results in MTT and 3H-TdR assays showed that growth inhibiting rate in the group of -45 ℃ was (3.83±0.56)% and (4.41±0.71)%, which was the lowest among the experimental groups (P < 0.05). -45 ℃ is the optimal frozen pretreatment temperature that can protect the bioactivity of embryonic rabbit’s Schwann cells. Related Articles | Metrics

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Isolation of murine muscle-derived stem cells with preplate technique combined with limited dilution technique Ding Wei-jin, Tang Yi, Song Yan-ling, Su Zhi-da, Li Cui, Liu An-tang, Hu Xia, Jiang Hua 2011, 15 (36):  6797-6801.  doi: 10.3969/j.issn.1673-8225.2011.36.034 Abstract ( 227 )   PDF (1519KB) ( 419 )   Save BACKGROUND: Muscle-derived stem cells (MDSCs), a newly discovered adult stem cell possessing utility potential in tissue engineering and regenerative medicine, have been isolated from skeletal muscle tissue. The MDSCs isolating method varies a lot. OBJECTIVE: To isolate and culture MDSCs and its clone as well as sub-clone through the use of modified preplate technique combined with limited dilution technique. METHODS: The muscle tissue was obtained from new born mice under microscopy and then digested and filtered, from which MDSCs were isolated by modified preplate technique with first round plating period of 1 hour. The muscle derived cells were counted and MDSCs were marked with immunohistochemistry method. The MDSC clone and its subclone were obtained with limited dilution technique. RESULTS AND CONCLUSION: During the isolation procedure with preplate technique, muscle derived cells made up a progressively higher ratio in the cell culture and the procedure with first round plating period of 1 hour provided plenty cells for MDSCs isolation. MDSCs presented with adherent growth 72 hours after the sixth suspension, grew into cell population of 300-500 cells in 10 days about, and proliferated with its small round and spindle morphology persisted. MDSCs clone and sub-clone were obtained through limited dilution technique and found desmin positively expressed and Sca-1 positively expressed at ratio of (92.3±4.1)%. The MDSCs and its clone from mice may provide proper cell resource for tissue engineering and regenerative medicine research. Related Articles | Metrics

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Influence of cerebrospinal fluid on in vitro culture of bone marrow mesenchymal stem cells Shen Yi-xin, Wang Peng, Shi En-dong 2011, 15 (36):  6802-6806.  doi: 10.3969/j.issn.1673-8225.2011.36.035 Abstract ( 206 )   PDF (648KB) ( 340 )   Save BACKGROUND: Cell transplantation is a promising strategy for treatment of spinal cord injury. But the current experimental evidence about cerebrospinal fluid (CSF) effect on the cells is lack. OBJECTIVE: To study the CSF effects on the in vitro culture of bone marrow mesenchymal stem cells (BMSCs). METHODS: BMSCs were isolated and purified by density gradient centrifugation and adherent culture, and BMSCs were passaged in vitro. The third and fourth passage cells were cultured in CSD to observe cells growth conditions, and analyze some of its phenotypic characteristics. The fifth passage cells were cultured in L-DMEM containing fetal calf serum and CSF. RESULTS AND CONCLUSION: We had successfully and effectively isolated and purified BMSCs by density gradient centrifugation with adherent culture. The results of the identification were positive of CD90, CD106, CD71, CD29, while CD45 was negative. Cells cultured in CSF still had still morphology of BMSCs. But few cells were neural-like cells. The results of the identification were positive of CD90, CD106, CD71, CD29, while CD45 was negative. Neuron-specific enolase was weakly positive. Cells had a similar S-shaped growth curve, and the growth curves were similar. BMSCs can still proliferate in CSF, thus it indicate that these cells possess high adaptability to their growth environment. Related Articles | Metrics

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Application of umbilical cord blood mesenchymal stem cells in cartilage tissue engineering Wang Cong-ren, Zhang Shou 2011, 15 (36):  6809-6812.  doi: 10.3969/j.issn.1673-8225.2011.36.037 Abstract ( 233 )   PDF (663KB) ( 318 )   Save BACKGROUND: Due to the poor repair capacity of articular cartilage, it is still a difficult problem to repair and reconstruct articular cartilage defects in orthopedics, and it is also one of the research foci in recent years. But, the development of cartilage tissue engineering provides a new way for repairing articular cartilage defects. OBJECTIVE: To conclude and analyze the biological characteristics of umbilical cord blood mesenchymal stem cells and their research and application in cartilage tissue engineering. METHODS: We searched CNKI and PubMed database for literatures from January 2000 to December 2010. Key words were “umbilical cord blood mesenchymal stem cells, tissue engineering, cartilage repair”. Articles on Umbilical cord blood mesenchymal stem cells and cartilage tissue engineering were included, and repetitive studies were excluded. Finally, 33 articles were selected for further analysis. RESULTS AND CONCLUSION: Umbilical cord blood mesenchymal stem cells, as ideal seed cells of cartilage tissue engineering, are easy to obtain, and have weak immunogenicity, strong differentiation, low rate of bacterial contamination by viruses and other unique advantages. And it is will play an important role in cartilage tissue engineering. Related Articles | Metrics

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Biological characteristics and application of epidermal stem cells Lai Qi, Xiong Ai-bing 2011, 15 (36):  6813-6816.  doi: 10.3969/j.issn.1673-8225.2011.36.038 Abstract ( 201 )   PDF (651KB) ( 272 )   Save BACKGROUND: To remove the necrotic tissue of the wound surfaces as early as possible, and rebuild the structure and function of skin is the ultimate therapeutic target of extensive deep burn. Epidermal stem cell as a specific stem cell in skin tissue has its incontestable potentials. OBJECTIVE: To review the research status on biological characteristics and application of epidermal stem cells in recent years. METHODS: An online search of PubMed database was performed using key words of “epidermal stem cells, basic study” for articles published between January 2000 and October 2010, and the language was limited to English. In the meantime, a computer search for Wanfang database was conducted using key words of “epidermal stem cells” for articles between January 2005 and October 2010, and the language was limited to Chinese. A total of 271 literatures were screened out, and finally 38 articles were included. RESULTS AND CONCLUSION: Epidermal stem cells are a kind of cells which have the potential of multi-directional differentiation, self-renewal and proliferation. Epidermal stem cells have the typical characteristics of long periodicity in vivo, capacity to easily adhere to skin basement membrane. The proliferation and differentiation of epidermal stem cells is regulated by Wnt signaling pathway,c-Myc proto-oncogene, and Notch signaling pathway. Currently, there are no specific markers of epidermal stem cells generally acknowledged. Epidermal stem cells can be explored in wound repair, gene therapy and tissue engineering. As the study getting intensive, epidermal stem cells will provide a new approach in the skin basic research, rehabilitation of wound function and skin heredopathia therapy. Related Articles | Metrics

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The research of periodontal ligament stem cells Zhang Ying, Song Li 2011, 15 (36):  6817-6820.  doi: 10.3969/j.issn.1673-8225.2011.36.039 Abstract ( 278 )   PDF (613KB) ( 416 )   Save BACKGROUND: Periodontal ligament stem cells isolated from periodontal ligament tissue is considered to be the preferred seed cells with self-renewal ability in periodontal tissue engineering and can differentiate to form three kinds of periodontal tissues: alveolar bone, periodontal ligament and cementum. OBJECTIVE: To review the articles about isolation, identification, and relevant factors of periodontal ligament stem cells in recent years. METHODS: The first author searched PubMed databases and CKNI database (2004-01/2010-09) for articles regarding isolation, identification, and relevant factors of periodontal ligament stem cells using the keywords of “periodontal ligament stem cells” in English and “periodontal ligament, stem cells” in Chinese. Repetitive articles were excluded, and finally, 35 articles were included. RESULTS AND CONCLUSION: Periodontal ligament stem cells are a kind of potential seed cells for periodontal tissue engineering, which can reconstruct tissue-engineered periodontal ligament and improve the healing of periodontal defects. Biological properties of periodontal ligament stem cells have been gradually found and their research remains to be further improved. Related Articles | Metrics

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Basic and clinical research of autologous bone marrow stem cells in treatment of hepatic cirrhosis Xing Ni-ni, Li Xiao-sheng 2011, 15 (36):  6821-6824.  doi: 10.3969/j.issn.1673-8225.2011.36.040 Abstract ( 341 )   PDF (675KB) ( 271 )   Save BACKGROUND: Liver transplantation is still facing many problems, such as lack of donor livers, immune rejection after transplantation and its high costs, thus making it restricted in clinic application. OBJECTIVE: To describe autologous bone marrow stem cell transplantation in the clinical stage as one of the most promising technology that autologous bone marrow stem cells can differentiate into mature liver cells to replace the damaged liver functioning, which can improve symptoms and quality of life in patients. METHODS: A computer-based online search of PubMed database, CNKI, Wanfang database and VIP database was performed for articles regarding autologous bone marrow stem cells to treat hepatic cirrhosis, with the key words of “bone marrow stem cell, autologous bone marrow stem cell transplantation, hepatic cirrhosis” in Chinese and “bone marrow stem cell, autologous bone marrow stem cell, hepatic cirrhosis” in English. Relevant articles published in recent years or in authorized journals were preferred. Repetitive studies were excluded. Finally, 20 articles were included. RESULTS AND CONCLUSION: Basic and clinical research about autologous bone marrow stem cell transplantation indicate that autologous bone marrow stem cell transplantation has a broad prospect in the treatment of hepatic cirrhosis. Related Articles | Metrics

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Tissue-engineered lung seed cells and extracellular matrix Feng Yi, Liu Sheng-ming 2011, 15 (36):  6825-6828.  doi: 10.3969/j.issn.1673-8225.2011.36.041 Abstract ( 270 )   PDF (610KB) ( 206 )   Save BACKGROUND: There is slow progress in the reconstruction and clinical application of tissue-engineered lung because of the difficulty in the identification and utilization of cell sources, development of standardized procedures for cell differentiation, control of extracellular matrix production to meet the needs of the lung and so on. OBJECTIVE: To review the identification and utilization of cell sources and the replacement of extracellular matrix in the tissue-engineered lung research. METHODS: A computer-based online search of China Journal Full-text Database and PubMed was performed for articles and reviews in Chinese and English from 1989 to 2011. The key words were “tissue-engineered lung, cell source, support scaffolding, lung matrices, vivo implantation”. Screened following reading titles and abstracts, relevant literatures were included. Repetitive articles with poor quality were excluded. A total of 72 articles were retrieved. Finally, 36 literatures were included. RESULTS AND CONCLUTION: New functional replacement lung tissue can be generated through tissue engineering in the use of cell sources and related support scaffolding under a certain condition, but it is still at the basic research stage. With the development of the techniques in the tissue-engineered lung, there will be more and more new discovery. The recent clinical use and success of decellularized trachea suggests that tissue-engineered lung may be the best choice for lung diseases in the future. Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cells transplantation on cardiomyocyte apoptosis related gene and protein expression Jiang Xiao-ming, Qu Hong-lin 2011, 15 (36):  6829-6832.  doi: 10.3969/j.issn.1673-8225.2011.36.042 Abstract ( 197 )   PDF (592KB) ( 242 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can act on myocardial cells, and prevent the occurrence of myocardial injury or secondary lesions. OBJECTIVE: To analyze the effects of bone marrow mesenchymal stem cells on cardiomyocyte apoptosis related gene and protein expression. METHODS: The first author searched PubMed and CNIKI databases (1997/2010) for articles regarding bone marrow mesenchymal stem cells, cardiomyocyte apoptosis and gene expression. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells become a hotspot because of its characteristics, such as easy to obtain, strong differentiation capacity, low immunogenicity, no rejection. Bone marrow mesenchymal stem cells can function in myocardial cells and prevent the occurrence of myocardial injury or secondary lesions, mainly through regulation of Bcl-2, Bax, Fas, FasL, Caspase gene and protein expression, to realize the inhibition of cardiomyocyte apoptosis. Bone marrow mesenchymal stem cells have a protective effect and applied value in the performance decline induced by cardiomyocyte apoptosis during the movement. Related Articles | Metrics

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Umbilical cord mesenchymal stem cell transplantation for treatment of Parkinson's disease in 8 cases Qiu Yun, Wang Zheng, Lu Hong-she 2011, 15 (36):  6833-6836.  doi: 10.3969/j.issn.1673-8225.2011.36.043 Abstract ( 307 )   PDF (541KB) ( 518 )   Save BACKGROUND: Neurons of the central nervous system neurons are a kind of terminal cells, and neuron loss caused by any pathological process is an irreversible process. OBJECTIVE: To observe the curative effect of umbilical cord mesenchymal stem cells (UC-MSCs) implantation in treating Parkinson’s disease (PD). METHODS: A total of 8 PD patients admitted in the Stem Cells Transplantation Center, the 455 Hospital of Chinese PLA, from August to December 2010, were selected, including 4 males and 4 females, aged 50-78 years. From the second week, the patients received UC-MSCs implantation by carotid artery puncturing. Unified Parkinson’s Disease Rating Scale (UPDRS) was used to evaluate those PD patients’ neural function. Higher scores indicate more severe neurological deficit. RESULTS AND CONCLUSION: All patients were included in the final analysis without loss. Compared to before transplantation, the UPDRS was significant decreased at 1 month after treatment (P < 0.01). The clinical symptoms, such as tremor, rigidity had obvious improvement. There was no significant improvement concerning the symptoms of slow movement and unstable posture. In addition, no graft versus host disease occurred. UC-MSCs implantation can ameliorate clinical symptoms, as well as improve life quality of PD patients to some extent. Related Articles | Metrics

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Umbilical cord blood mesenchymal stem cell transplantation for treatment of a child with spinal muscular atrophy Du Ling, Yang Hua-qiang, Wang Na, Luo Guo-jun 2011, 15 (36):  6837-6840.  doi: 10.3969/j.issn.1673-8225.2011.36.044 Abstract ( 323 )   PDF (287KB) ( 432 )   Save BACKGROUND: Many animal and clinical studies have reported that the safe and effective usage of umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) transplantation for treatment of neurological genetic diseases. OBJECTIVE: To investigate the feasibility and effect of UCB-MSCs transplantation in the treatment of spinal muscular atrophy (SMA). METHODS: A child admitted at January 2010 had been confirmed as having SMA, and drug and rehabilitation therapies were invalid. Then, the child received UCB-MSCs transplantation via the first intravenous infusion and three times of subarachnoid injection, once a week, (4-6)×107 cells once and four times as a course. Neurological physical examination, biochemical test, muscle enzymes detection, FIM scoring and electromyography (EMG) examination were conducted. RESULTS AND CONCLUSION: Compared with prior to transplantation, the level of muscle enzymes decreased, FIM scores were increased from 68 to 93 points, EMG results showed that the motor units with re-contraction in each 10.0 ms were increased that the motor function was improved, the lower extremity muscle strength elevated, and the self-care ability was improved in the SMA child at 6 months after transplantation. During the 10-month follow-up, the child had no adverse effects. It is indicated that UCB-MSCs transplantation is effective to treat SMA, and the neurological function has a remarkable restoration. Related Articles | Metrics