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06 August 2011, Volume 15 Issue 32 Previous Issue    Next Issue

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Modalities of the first medium exchange during the culture of rat bone-marrow derived mesenchymal stem cells Ni Ping, Chen Zi-qian, Peng De-xin, Bai Wei 2011, 15 (32):  5891-5895.  doi: 10.3969/j.issn.1673-8225.2011.32.001 Abstract ( 385 )   PDF (1419KB) ( 519 )   Save BACKGROUND: The culture period, quantity and purity are crucial for stem cell transplantation, and it is essential to build an optimal modality of cell culture in vitro.  OBJECTIVE: To study the influence of different modalities of the first medium exchange on the culture time and biological characteristics of rat bone marrow-derived mesenchymal stem cells. METHODS: On condition of modality of the first medium exchange, six experimental groups were performed as follows: group A was half exchanged after 24 hours, group B was all exchanged after 24 hours, group C was half exchanged after 48 hours, group D was all exchanged after 48 hours, group E was half exchanged after 72 hours, group F was all exchanged after 72 hours. Then, all groups were all exchanged every 48 hours and were subcultured consecutively. The period and purity of each passage were measured. RESULTS AND CONCLUSION: The total culture time was significant different (F=46.455, P < 0.001). Group D was the least while group B was the longest, and both of the two groups were significant different compared with others (P < 0.05). Group B earned the highest purity in all passages and group E the lowest, and both of the two groups were significant different compared with others (P < 0.05). Cells of passage 4 showed CD29, CD90 positive, induced to differentiate into osteoblasts and adipocytes. The modality of the first medium exchange had an influence on the culture period and purity of bone marrow-derived mesenchymal stem cells, and the modality of 48-hour half exchange was the best protocol. Related Articles | Metrics

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Improvement of methods to isolate and culture human mesenchymal stem cells Wang Cheng-yun, Shi Lei, Xia Chun, Zheng Xin-peng, Zhang Bing, Lin Fei-tai, Wang Shao-jie 2011, 15 (32):  5896-5900.  doi: 10.3969/j.issn.1673-8225.2011.32.002 Abstract ( 328 )   PDF (1572KB) ( 474 )   Save BACKGROUND: The methods to isolate mesenchymal stem cells (MSCs) can be classified to be several kinds. However, the method of acquisition in surgery is still less. The isolated MSCs from the patients who receive the replacement operation can offer some data which may reveal the causes or clues of some diseases. OBJECTIVE: To isolate and culture MSCs with an improved technique of differential anchoring velocity and adherence, in order to identify MSCs. METHODS: The bone marrow blood was collected in the femur marrow cavity duiring the hip joint replacement. MSCs were isolated by adherence method in the special medium and the mix medium was flushed 36-48 hours after the inoculation to isolate MSCs from the marrow mixture. In the long-time culture in vitro, the special medium was used to weed other cells out. The anchored cell was cultured and identified when the amount was large enough.  RESULTS AND CONCLUSION: MSCs can be isolated from the bone marrow in long cavitas medullaris, and can be cultured into colonies by using adherence method. The morphological features and surface molecules are identified consistent with MSCs, and can obtain standard MSCs, so as to do the follow-up experiment. Related Articles | Metrics

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Immunoregulatory function of fetal bone marrow-derived mesenchymal stem cells on human Th17 cells Guo Zhen-xing, Zheng Cui-ling, Chen Zhen-ping, Dong Wen-chuan, Yang Ren-chi 2011, 15 (32):  5901-5904.  doi: 10.3969/j.issn.1673-8225.2011.32.003 Abstract ( 256 )   PDF (1100KB) ( 348 )   Save BACKGROUND: Many studies have demonstrated mesenchymal stem cell (MSC) can regulate the immune function and inhibit proliferation of T cell. OBJECTIVE: To investigate the regulatory function of fetal bone marrow-derived mesenchymal stem cells (FBM-MSCs) on human Th17 cells. METHODS: FBM-MSCs were cocultured with human peripheral blood mononuclear cells (PBMCs) or CD4+ T cells at 1:10 ratio from healthy donors for 4 days. Single culture of mononuclear cells or CD4+ T cells were as controls. The expression of interleukin-17 (IL-17) mRNA was detected by real-time PCR, IL-17 protein was assayed by enzyme-labeled immunosorbent assay (ELISA), and quantity of Th17 cells was observed by flow cytometry. RESULTS AND CONCLUSION: The level of IL-17mRNA in FBM-MSC cocultured with PBMC (FBM-MSC/PBMC) group was significantly higher than that in PBMC cultured alone. Meanwhile, IL-17 expression in supernatant in FBM-MSC/PBMC/ CD4+ was significantly higher than that in PBMC and CD4+ T cells groups ( P < 0.05, P < 0.01). The number of FBM-MSCs/CD4+ T cells group was significantly more than CD4+ T cells group ( P < 0.01). However, FBM-MSCs did not express IL-17 protein. FBM-MSCs can promote the proliferation of Th17 cells. Related Articles | Metrics

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Effect of DNA demethylation on proliferating cell nuclear antigen expression in bone marrow mesenchymal stem cells Zhao Hong-xian, Xu Fu-cui, Wang Qiao-zhi, Xu Qiang, Chen Xia 2011, 15 (32):  5905-5908.  doi: 10.3969/j.issn.1673-8225.2011.32.004 Abstract ( 248 )   PDF (1119KB) ( 343 )   Save BACKGROUND: DNA demethylation is an important epigenetic modification. OBJECTIVE: To explore the effect of DNA demethylation on proliferation and proliferating cell nuclear antigen (PCNA) protein 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, CD44, CD45 and PCNA protein were dectcted by immunocytochemistry, and effect of DNA demethylation on proliferation was detected by MTT method. RESULTS AND CONCLUSION: ①The third passage bone marrow mesenchymal stem cells expressed CD44 postively and CD45 negatively. ②Compared with control group, after 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased proliferation of cells without concentration-dependence significantly (P < 0.05). ③ Compared with control group, after 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased PCNA protein expression significantly in bone marrow mesenchymal stem cells without concentration-dependence (P < 0.05). In certain concerntration, 5-azacytidine can increase PCNA protein expression thirough DNA demethylation, which is the key to improve proliferation of bone marrow mesenchymal stem cells. Related Articles | Metrics

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Differentiation of bone marrow mesenchymal stem cells into thyroid cells Xin Qun, Liu Dong-yuan, Zhang Qin 2011, 15 (32):  5909-5912.  doi: 10.3969/j.issn.1673-8225.2011.32.005 Abstract ( 348 )   PDF (1067KB) ( 319 )   Save BACKGROUND: Under certain conditions, rat bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into thyroid cells. OBJECTIVE: To establish the method to in vitro induce the differentiation of rat BMSCs into thyroid cells. METHODS: BMSCs were separated using density gradient method, and induced with thyroid-stimulating hormone and insulin. A group without any induced materials was used as parallel control. Morphology changes were observed with inverted optical microscope and transmission electron microscopy, immune fluorescence detection method was applied for BMSCs differentiation. RESULTS AND CONCLUSION: Thyroid cell-specific gene expression such as TSHr could be seen in cultured cells after 7 days; thyroid cell-differentiated cell markers TTF-1 expression was detected after 9 days; no change in the control group. BMSCs can be induced to differentiate into thyroid cells based on morphology and biological examination. Related Articles | Metrics

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Effects of simvastatin on bone mass and bone marrow mesenchymal stem cells proliferation in rats Liang Chun-yu, Tian Fa-ming, Zhang Hui, Zhang Liu, Zhang Ying-ze 2011, 15 (32):  5913-5917.  doi: 10.3969/j.issn.1673-8225.2011.32.006 Abstract ( 321 )   PDF (1507KB) ( 314 )   Save BACKGROUND: As a widely used lipid-lowering drug, simvastatin has been rarely reported addressing its effects on bone metabolism and proliferation and differentiation of bone marrow stromal cells (BMSCs).   OBJECTIVE: To investigate the effects of simvastatin on bone mass and differentiation and proliferation of BMSCs in rats, as well as the expression levels of Smad1, 2, 7 mRNA. METHODS: Sixteen 6-week-old female Sprague-Dawley rats were randomized into two groups of eight animals each: simvastatin group, administered daily with 20 mg/kg simvastatin by gavage for 9 weeks; control group, administered with distilled water for 9 weeks. All animals were sacrificed one day after the final administration. The left femora were removed for the measurement of bone histomorphometry and bone mineral density (BMD). BMSCs derived from the right femora and tibiae were cultured in osteoblastic differentiation-induced medium. Cell Counting Kit-8 (CCK-8) was used to assay the proliferation of BMSCs. Alkaline phosphatase (ALP) activity, ALP staining were performed on the 16th day and von Kossa staining on the 28th day. Real-time RT-PCR was used to evaluate the expression levels of mRNA of Smad1, 2, 7 at the 21st day. RESULTS AND CONCLUSION: After being administrated with simvastatin for 9 weeks, the bone mass and BMD of rats had no significant change. The expression level of mRNA of Smad1, 2, 7 showed no significant change, so were the results of ALP activity, positive cell rate of ALP staining, von Kossa staining, and the proliferation of BMSCs. Administration with 20 mg/kg simvastatin for 9 weeks had no significant effect on bone mass and the differentiation and proliferation of BMSCs in rats, so were the expression levels of Smad1, 2, 7 mRNA. Related Articles | Metrics

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Ultrastructural changes following osteogenic differentiation of rat bone marrow mesenchymal stem cells Pan Sheng-cai, Tang Yu-jin, Xie Ke-gong, Lan Chang-gong, Lu Xian-zhe, Li Xiao-feng 2011, 15 (32):  5918-5922.  doi: 10.3969/j.issn.1673-8225.2011.32.007 Abstract ( 244 )   PDF (1539KB) ( 439 )   Save BACKGROUND: There are few studies on ultrastructure characteristics of osteoblasts after differentiation of bone marrow mesenchymal stem cells (BMSCs) OBJECTIVE: By using the whole bone marrow adherence method, to isolate, culture, purified BMSCs from rats, and to observe the ultrastructural changes after osteogenic differentiation of rat BMSCs. METHODS: BMSCs from rats were isolated, cultured and purified by the whole bone marrow adherence method; for morphology observations, cell surface markers were assessed by flow cytometry, and BMSCs were induced to differentiate into osteoblasts and stained to identification. Scanning electron microscope and transmission electron microscope were used to observe the ultrastructural changes of cells before and after induction. RESULTS AND CONCLUSION: The third generation cultured BMSCs had high purity and strong vitality. After induction, BMSCs were positive in bone alkaline phosphatase activity and calcified nodules staining. Scanning electron microscope and transmission electron microscope observation showed that after osteogenic induction, the cells spread, arranged irregularly, and were rich of organelles; and the cell mitochondria, rough endoplasmic reticulum, vacuoles increased significantly, indicating that the cells were functionally active. Related Articles | Metrics

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Culture condition and biological characteristics of rat bone marrow mesenchymal stem cells by using the whole bone marrow adherence method Zhao Lin, Feng Zhi-hui, Jiao Shu-xian, Li Nan-nan 2011, 15 (32):  5923-5927.  doi: 10.3969/j.issn.1673-8225.2011.32.008 Abstract ( 300 )   PDF (1413KB) ( 709 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used as seed cells in tissue engineering and other fields. Therefore, well growth and enough number of rat BMSCs are important for above-mention studies. OBJECTIVE: To explore the suitable primary culture condition of rat BMSCs in vitro, and to observe the growth property of BMSCs. METHODS: BMSCs were separated using whole bone marrow culture method and purified using attachment method. Morphology and growth characteristics were observed under inverted phase contrast microscope and cytoskeleton was detected by immunofluorescence. Growth curve was drawn using MTT method. Expression of BMSC surface antigens was detected by flow cytometer. RESULTS AND CONCLUSION: After isolation and culture, cells developed a spindle or polygonal appearance. Growth curve was S-shaped, and the cultured BMSCs possessed normal morphology and cytoskeleton. At the third passage, BMSCs were positive for CD29, CD44, CD71, CD90, but negative for CD13, CD34, CD45, CD133. Cells isolated and cultured by the adherence separation method in this study are identified as rat BMSCs by morphology and surface antigens, which have the biological characteristics of BMSCs. Related Articles | Metrics

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Comparison of bone marrow-derived mesenchymal stem cells from aborted fetus and normal adult subjects Cheng Yue-ai, Liu Rui-feng, Zhang Yong-cui, Zhang Kai-ming 2011, 15 (32):  5928-5931.  doi: 10.3969/j.issn.1673-8225.2011.32.009 Abstract ( 251 )   PDF (1269KB) ( 327 )   Save BACKGROUND: Bone marrow-derived mesenchymal stem cells are pluripotent cells and can be involved in immune response. Fetal and adult bone marrow is both important sources for mesenchymal stem cells. OBJECTIVE: To reveal the differences of bone marrow derived mesenchymal stem cells from aborted fetus and normal adult subjects by comparing the biological characters (including morphology, immunophenotype, proliferative activity and cytokines secretion). METHODS: Bone marrow mononuclear cells of aborted fetus and normal adult subjects were separated by density gradient centrifugation. Bone marrow derived mesenchymal stem cells were cultured with the adherent method. Cells at passage 3 and their culture supernatants were collected. Flow cytometry analysis was used to examine the immunophenotype. The proliferation activity of cells was measured by MTT colorimetric assay. ELISA was used to measure the concentrations of cytokines secreted from bone marrow derived mesenchymal stem cells in the two groups.  RESULTS AND CONCLUSION: Cells from both groups have the same morphology, immunophenotype and the levels of cytokines secretion. But in comparison with normal adult subjects, bone marrow derived mesenchymal stem cells from aborted fetus showed higher proliferative activity.  Bone marrow derived mesenchymal stem cells from aborted fetus and normal adult subjects have the similar biological characteristics except the proliferative activity. Aborted fetus can provide bone marrow for research of stem cells. Related Articles | Metrics

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In vitro differentiation capacity of adipose-derived stromal stem cells and bone marrow stromal stem cells Zhou Jin, Tian Guo-ping, Wang Jing-e, Xu Bing, Li Li, Zhu Feng, Han Jian, Hu Feng-jie 2011, 15 (32):  5932-5935.  doi: 10.3969/j.issn.1673-8225.2011.32.010 Abstract ( 421 )   PDF (572KB) ( 354 )   Save BACKGROUND: Adipose-derived stromal stem cells (ADSCs) have many biological properties similar to bone marrow stromal stem cells (BMSCs). OBJECTIVE: To compare the directed differentiation of ADSCs and BMSCs co-cultured with damaged PC12 cells. METHODS: Stromal stem cells were isolated from adipose tissue and bone marrow. The fifth passage cells were selected. Two kinds of cells were co-cultured, respectively, with or without normal or damaged PC12 cells supernatant. RESULTS AND CONCLUSION: CD44 and CD29 highly expressed in ADSCs and BMSCs, while CD45 and CD56 were only detected in BMSCs, but rare in ADSCs. After single culture, two kinds of cells highly expressed Nanog, Oct4, Sox2, but not neuron-specific enolase (NSE). The levels of Nanog, Oct4, Sox2 in the cells co-cultured with damaged PC12 cells were significantly lowered, while NSE positive cells were increased in ADSCs. These indicate that damaged PC12 cells may have a stronger effect on induce and differentiation of ADSCs.  Related Articles | Metrics

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Comparison of the biological characteristics of bone marrow and adipose tissue derived mesenchymal stem cells Zhu Xi-shan, Shi Wei, Tai Wei-ping, An Guang-yu 2011, 15 (32):  5936-5940.  doi: 10.3969/j.issn.1673-8225.2011.32.011 Abstract ( 313 )   PDF (596KB) ( 439 )   Save BACKGROUND: Recently, adipose tissue derived mesenchymal stem cells due to easy to be harvested have been widely studied. OBJECTIVE: To compare the biological characteristics of adipose tissue derived mesenchymal stem cells and bone marrow mesenchymal stem cells. METHODS: Adipose tissue derived mesenchymal stem cells and bone marrow mesenchymal stem cells were isolated and cultured in vitro to compare their phenotype, cell doubling time and the level of secreted factors. RESULTS AND CONCLUSION: Adipose tissue derived mesenchymal stem cells have similar phenotype and differentiation ability as bone marrow mesenchymal stem cells except CD106 expression. The frequency of mesenchymal stem cells in bone marrow and adipose tissue was also calculated and found that mesenchymal stem cells frequency in adipose tissue was 10 times higher than bone marrow. This result can be an instruction for mesenchymal stem cells clinical use. Related Articles | Metrics

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Neuroprotective effect of mild hypothermia on the proliferation of endogenous neural stem cells in rats with intracerebral hemorrhage Zhang Jun, Zhu Ping, Wang Lian-kun, Hou Dan-hui, Wang Ying, Wang Kun-xiang 2011, 15 (32):  5941-5944.  doi: 10.3969/j.issn.1673-8225.2011.32.012 Abstract ( 242 )   PDF (694KB) ( 347 )   Save BACKGROUND: There are a few reports addressing application of mild hypothermia to brain edema after intracerebral hemorrhage, but few reports have addressed mild hypothermia effects on proliferation of neural stem cells in the brain following intracerebral hemorrhage. OBJECTIVE: To explore the effects of mild hypothermia on proliferation of neural stem cells of rats after intracerebral hemorrhage. METHODS: A model of intracerebral hemorrhage was established by autologous arterial blood injected into right caudate nucleus. Local mild hypothermia was applied in the mild hypothermia group for 4 hours after the blood injected in the brain. Normal body temperature was maintained in control group. RESULTS AND CONCLUSION: At 1, 3, 7, 14 days after intracerebral hemorrhage, the Longa score in the hypothermia group was lower than that in the control group (P < 0.05). At various time points following intracerebral hemorrhage, the number of BrdU-positive cells was obviously greater in the hypothermia group compared with the control group (P  < 0.05). Results had indicated that mild hypothermia can promote the proliferation of neural stem cells, and shows neuroprotective effects on intracerebral hemorrhage. Related Articles | Metrics

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Remnant-like particles activate peroxisome proliferator-activated receptor gamma and induce adipogenic differentiation of human adipose mesenchymal stem cells Ma Mei-fang, Wen Tie, Liu Ling 2011, 15 (32):  5945-5950.  doi: 10.3969/j.issn.1673-8225.2011.32.013 Abstract ( 365 )   PDF (1832KB) ( 507 )   Save BACKGROUND: Remnant-like particles (RLPs) can induce rat adipose-derived mesenchymal stem cells (ADSCs) to differentiate into mature adipocytes. OBJECTIVE: To explore the effects of RLPs on the adipogenic differentiation of human ADSCs and on mRNA expressions of peroxisome proliferator-activated receptor γ (PPAR-γ) and adiponectin (APN). METHODS: After stimulation by standard cocktail inducer, lipid droplets within adipocytes were identified by oil red O staining. Human RLPs were isolated by density gradient ultracentrifugation and immuno-affinity chromatography from postprandial plasma of hypertriglyceridemic patients at 4 hours after a high-fat meal. ADSCs were stimulated by 10 mg/L insulin and different concentrations of RLPs (0, 50, 100, 150, 200 mg/L).  RESULTS AND CONCLUSION: After stimulation at 12 days, a small amount of lipid droplets was found in the control group or  50 mg/L RLPs stimulated ADSCs by oil red O staining. Lipid droplets increased gradually with the increase of RLPs concentrations from 100 to 200 mg/L and reached the peak in 200 mg/L RLPs stimulated ADSCs. The mRNA expressions of PPAR-γ and APN increased with the increase of RLPs concentrations from 50-200 mg/L (P < 0.05). APN mRNA expression in 200 mg/L RLPs group was significantly weaker than that in 150 mg/L RLPs group (P < 0.05), while it showed no significant difference when compared with that in 100 mg/L RLPs group. It indicates that RLPs induce adipogenic differentiation of human ADSCs through activating PPARγ, and up-regulate APN expression. Related Articles | Metrics

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Differentiation potential of human Orbicularis oculi muscle-derived stem cells towards Schwann cells phenotype Ding Wei-jin, Zhang Wen-jun, Sun Mei-qing, Su Zhi-da, Li Cui, Liu An-tang, Jiang Hua 2011, 15 (32):  5951-5956.  doi: 10.3969/j.issn.1673-8225.2011.32.014 Abstract ( 257 )   PDF (1913KB) ( 402 )   Save BACKGROUND: Muscle derived stem cells (MDSCs) can be isolated from human orbicularis oculi muscle and be differentiated to a Schwann cell phenotype which could eventually provide functional benefits for peripheral nerve repair. OBJECTIVE: To induce the differentiation of MDSCs into Schwann cell phenotype. METHODS: ①Under the support of microscope, we collected the discarded human Orbicularis oculi muscle resected in the upper eyelid blepharoplasty and isolate human-MDSCs within it with aid of tri-enzyme digestion and pre-plating technique, and then identify the cells by immunohistochemistry method. ②We isolated Schwann cells and identify the cells by immunohistochemistry method. Through half-harvest method, we would like to prepare conditioned medium from Schwann cell culture. ③We co-culture human-MDSCs with Schwann cell conditioned medium and the transdifferentiated cell morphology was investigated daily under microscope. The common used marker, S-100, GFAP and p75 were stained to identify Schwann cell phenotype with use of immunohistochemistry method. RESULTS AND CONCLUSION: ①We collected human Orbicularis oculi muscle sample from three young female volunteer with their consensus. Human-MDSCs were isolated from Orbicularis oculi muscle and have their desmin positively stained and Sca-1 was positively expressed. ②Schwann cells were isolated and identified with S-100 positively stained at the rate of (97.4±0.7)%. ③The isolated human-MDSCs were successfully transdifferentiated into Schwann cell-like cells with positive expression of S100, GFAP and p75, which would serve as a unanimous evidence of Schwann cell phenotype. Human-MDSCs could be transdifferentiated into Schwann cell-like cells when co-cultured within Schwann cell conditioned medium, which would serve as an alternative candidate for commonly studied Schwann cells in tissue engineering nerve graft. Related Articles | Metrics

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Neuregulin-1 induces embryonic stem cells to differentiate into cardiomyocytes and its mechanism Wang Zhi, Huang Jin 2011, 15 (32):  5957-5961.  doi: 10.3969/j.issn.1673-8225.2011.32.015 Abstract ( 300 )   PDF (1732KB) ( 386 )   Save BACKGROUND: Previous studies demonstrated that the early invention of neuregulin-1 (NRG-1) can induce embryonic stem cells (ESCs) to differentiate into cardiomyocytes. OBJECTIVE: To further investigate the pathway of NRG-1 induces embryonic stem cells to differentiate into cardiomyocytes. METHODS: The process of cardiomyocytes (CMs) differentiated from ESCs with or without NRG-1 treatment was observed continuously. RT-PCR was performed to detect the mRNA expression of early cardiac-restricted transcript factors Nkx2.5 and GATA-4 during differentiation. The protein expression of cardiac troponin T and phosphorylated protein kinase B was determined by immunhistochemistry (IHC), phosphorylated protein kinase B expression of CMs was analyzed by semiquantitative western blot analysis. RESULTS AND CONCLUSION: The differentiation rate of CMs induced by NRG-1 was significantly higher than spontaneous CMs differentiation rate. NRG-1 exhibited a dose dependent up regulatory effect on the expression of GATA-4, Nkx2.5 mRNA, while NRG-1-induced increase of Nkx2.5 transcripts was inhibited by treatment with receptor antagonist and phosphatidylinositol 3-kinase (PI3K) inhibitor. Western blot analysis confirmed that the expression of phosphorylated protein kinase B of spontaneous beating CMs in embryoid body in NRG-1 induced group was relatively higher than that in spontaneous CMs differentiation group. The results suggested that NRG-1 promotes CMs differentiation of mouse ESCs via PI3K/phosphorylated protein kinase B/ phosphorylated protein kinase B activation. Related Articles | Metrics

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Effects of edaravone on endogenous neural stem cells in cerebral ischemic rats Han Qiu, Shen Li-hua, Zhang Cui, Wu Er-bing, Liu Xiao-fei 2011, 15 (32):  5962-5966.  doi: 10.3969/j.issn.1673-8225.2011.32.016 Abstract ( 322 )   PDF (1626KB) ( 410 )   Save BACKGROUND: Edaravone (MCI-186) is a novel free radical scavenger. Many studies have confirmed that MCI-186 can inhabit brain edema in acute cerebral infarction and produce neuroprotective effects. OBJECTIVE: To investigate the effect of free radical scavenger MCI-186 on endogenous neural stem cells around ischemic brain regions after cerebral infarction. METHODS: The middle cerebral artery ischemia and reperfusion model of SD rats was established with Longa method and divided into two groups. They were administered with MCI-186 or phosphate buffered solution (PBS) immediately after artery occlusion. The contents of malondialdehyde and brain derived neurotrophic factor expressions around ischemic brain regions were monitored at 1, 3 and 7 days chronologically. Immunohistochemistry was used to monitor the expression of Nestin and Caspase-3 positive cells around the ischemic area. Neurological function was evaluated at the same time. RESULTS AND CONCLUSION: Compared with sham-surgery group, the malondialdehyde content in PBS group was increased, but reduced in the MCI-186 group (P < 0.01). The levels of brain derived neurotrophic factor protein and mRNA were both significantly increased in the PBS group at day 1 after ischemia (P < 0.01). MCI-186 enhanced the secretions and prolonged high level to day 3 compared with sham group and PBS group (P < 0.01). At days 3 and 7, the number of Nestin-positive cells in ischemic brain in MCI-186 group was notably higher than that in PBS group (P < 0.05), while Caspase-3 positive cells decreased (P < 0.05). The neurological function was obviously improved in MCI-186 group than that in PBS group at day 7. MCI-186 can inhibit lipid peroxidation, increase the secretion of brain derived neurotrophic factor in ischemic brain, protect neural stem cells from apoptosis, and confer neuroprotective effects. Related Articles | Metrics

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Effects of bone morphogenetic proteins 2 on neural stem cells of 14-day-old fetal rat telencephalon differentiating into cholinergic neurons Liang Jun, Li Ming-qiu, Zhao Fu-sheng, Dong Jian-jiang, Li Yue-zhen 2011, 15 (32):  5967-5970.  doi: 10.3969/j.issn.1673-8225.2011.32.017 Abstract ( 236 )   PDF (1098KB) ( 404 )   Save BACKGROUND: Bone morphogenetic protein-2 (BMP2) may be an extracellular regulatory factor involved in cholinergic differentiation of neuronal precursor cells. OBJECTIVE: To explore the effects of BMP2 on neural stem cells of 14-day-old fetus rat telencephalons induced into cholinergic neurons. METHODS: Fetus telencephalons of 14-day-old SD rats were isolated, and tissues were divisively treated with collagenase typeⅠcontained EDTA following trituration. The cells were raised in polylysine culture plates with serum-free medium. Primary culture medium was changed half after 24 hours, and 10 μg/L BMP2 was added. RESULTS AND CONCLUSION: The adherent monoculture neural stem cells could been obtained using collagenase. The Nestin positve cells were obtained, and the purity was over 99%. After induced by BMP2, the ChAT positve cells were obtained, and the purity was over 97%. BMP2 can induce neural stem cells of 14-day-old fetus rat telencephalons into cholinergic neurons. Related Articles | Metrics

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Bone morphogenetic protein 9 induces skeletal muscle-derived stem cells differentiating into osteoblasts Li Xiang, Lin Chun-yang, Qiu Bei-ming, Chen Liang, Deng Zhong-liang 2011, 15 (32):  5971-5974.  doi: 10.3969/j.issn.1673-8225.2011.32.018 Abstract ( 254 )   PDF (1202KB) ( 435 )   Save BACKGROUND: Previous studies found that some bone morphogenetic proteins can induce mesenchymal stem cells to differentiate into osteoblasts. OBJECTIVE: To assess the capacity of bone morphogenetic protein 9 to induce the differentiation of skeletal muscle-derived stem cells into osteoblasts. METHODS: Rabbit skeletal muscle-derived stem cells were obtained by the two-step method of collagenase-Ⅰ and trypsin, and transfected by bone morphogenetic protein 9 through recombinant adenoviruses. Then, the effect of bone morphogenetic protein 9 on the differentiation of skeletal muscle-derived stem cells into osteoblasts was observed by testing the alkaline phosphatase quantitation and calcium deposition. RESULTS AND CONCLUSION: Muscle-derived stem cells were successfully isolated from the rabbit and over 80% of muscle-derived stem cells showed Sca-1 positive immunoreactivity. Bone morphogenetic protein 9 induced alkaline phosphatase activities in skeletal muscle-derived stem cells and promoted calcium deposition of the cells. From the above, bone morphogenetic protein 9 has the capability to induce the differentiation of skeletal muscle-derived stem cells into osteoblasts． Related Articles | Metrics

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Detection of pluripotent transcription factors in fetal dermis-derived mesenchymal stem cells Yang Shao-guang, Xing Wen, Tian Kun, Liu Meng, Lu Shi-hong, Zhao Qin-jun, Ren Hong-ying, Pang Ai-ming, Chi Ying, Ma Feng-xia, Han Zhong-chao 2011, 15 (32):  5975-5978.  doi: 10.3969/j.issn.1673-8225.2011.32.019 Abstract ( 348 )   PDF (1163KB) ( 384 )   Save BACKGROUND: The induced pluripotent stem (iPS) cells have been reprogrammed from human dermal fibroblasts and mesenchymal stem cells (MSCs). Various combinations of transcription factors are used by different investigators. OBJECTIVE: To detect the pluripotent transcription factors expressed in fetal dermis-derived MSCs.  METHODS: Using previous methodology in isolation and cultivation of MSCs, fetal dermis-derived MSCs were obtained from 5-month-old fetus, inducing abortion by water bag. RESULTS AND CONCLUSION: Transcription factors of Oct4 and C-myc were expressed highly and Sox2 was expressed moderately. These results suggest that fetal dermis-derived MSCs are better somatic cells for iPS cells. Related Articles | Metrics

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Bionomics of mesenchymal stem cells from Wharton’s Jelly of the human umbilical cord Li Xiao-ling, Zhu Lü-yun, Zhao Fang 2011, 15 (32):  5979-5982.  doi: 10.3969/j.issn.1673-8225.2011.32.020 Abstract ( 372 )   PDF (1228KB) ( 643 )   Save BACKGROUND: Mesenchymal stem cells from Wharton’s Jelly of the human umbilical cord present with low immunogenicity and better proliferative capacity than bone marrow mesenchymal stem cells, which attract widespread attention. OBJECTIVE: To isolate and culture mesenchymal stem cells from Wharton’s Jelly of the human umbilical cord and to study their bionomics. METHODS: Mesenchymal stem cells were isolated and cultured from Wharton’s Jelly of the human umbilical cord by tissue cultivation. The growth curve was determined by MTT. Immunophenotype was identified by flow cytometry after dissociation. RESULTS AND CONCLUSION: Under inversed microscope, stem cells crawled from the human umbilical cord on the second day. About seven days later, fusiform cells appeared. The shape of cells was uniform in passage 3. There was no significant change on the shape of cells in passage 7 and passage 10. The cells were still to remain fusiform.The result of MTT showed that double time of UC-MSCs in passage 3 was 3 d to 4 d. There was no significant change in the shape of passages 7 and 10 cells. UC-MSCs expressed CD44, CD29, CD105. Cells were negative for the lineage markers of CD34, CD45, CD14. Related Articles | Metrics

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Wound healing related trophic factors derived from human umbilical cord mesenchymal stem cells Zhang Mei-rong, Sun Ya-ru, Wang Li, Zhang Xue-feng, Hu Jian-xia, Zhou Qian, Gao Hong, Wang Yan-gang 2011, 15 (32):  5983-5986.  doi: 10.3969/j.issn.1673-8225.2011.32.021 Abstract ( 339 )   PDF (1257KB) ( 483 )   Save BACKGROUND: Few reports have quantitatively determined the levels of wound healing related factors in the conditioned media of human umbilical cord mesenchymal stem cells (HUMSCs). OBJECTIVE: To measure the secretion levels of interleukin-6 (IL-6), IL-8, transforming growth factor 1 (TGF-1), monocyte chemoattractant protein 1 (MCP-1), vascular endothelial growth factor (VEGF), GM-CSF and TIMP-1 in HUMSCs derived conditioned media of 10 different individuals. METHODS: For culturing HUMSCs, CCS1107 was used as fetal bovine serum supplement to the culture medium. 1.1x104/cm2 P2 cells were seeded in 100 mm diameter dishes, and after 96 hours’ culture, the supernatant was collected and applied to ELISA measurement. RESULTS AND CONCLUSION: No significant secretion difference among 10 different individuals and between genders was observed. Of total factors being measured, the levels of MCP-1 and TIMP-1 were the highest. These results indicated that HUMSCs derived conditioned media contains numerous kinds of wound healing related factors, which may clinically provide a novel therapeutic method for treating various wound healing related impairments. Related Articles | Metrics

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Effect and safety of umbilical cord mesenchymal stem cells transplantation on end-stage liver cirrhosis Zhou Bing-xi, Guo Jun-xian, Han Shuang-yin, Zhang Yan-rui 2011, 15 (32):  5987-5990.  doi: 10.3969/j.issn.1673-8225.2011.32.022 Abstract ( 394 )   PDF (1456KB) ( 682 )   Save BACKGROUND: Bone mesenchymal stem cells (BMSCs) transplantation has been used for end-stage liver cirrhosis, and obtained some results. OBJECTIVE: To observe the effect and safety of umbilical cord mesenchymal stem cells (UCMSCs) transplantation on end-stage hepatic cirrhosis. METHODS: UCMSCs were isolated under sterile conditions. Sixty cases of liver cirrhosis received basic medication and UCMSCs transplantation by way of hepatic artery. RESULTS AND CONCLUSION: After UCMSCs treatment, the level of albumin, prealbumin and fibrinogen were significantly increased (P < 0.05), and the levels of total bilirubin and prothrombin time were obviously reduced (P < 0.05). The symptoms such as fatigue, abdominal distension, anorexia relieved after treatment. UCMSCs transplantation for end-stage liver cirrhosis is safe and effective, and it can improve patients’ liver function, blood clotting and relieve clinical symptoms. Related Articles | Metrics

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Effects of intravenous transplantation of umbilical blood mesenchymal stem cells on cerebral edema secondary to hematoma in neonatal rats Yan Xiao-hua, Chen Qi-wen, Xu Xin, Li Yan-cheng, Yan Shao-ming, Huang Zhen-gang, Zhang Li-na 2011, 15 (32):  5991-5994.  doi: 10.3969/j.issn.1673-8225.2011.32.023 Abstract ( 315 )   PDF (1307KB) ( 306 )   Save BACKGROUND: There are few reports about human umbilical cord blood mesenchymal stem cells (HUCBMSCs) for cerebral edema secondary to hematoma in neonatal animals. OBJECTIVE: To study the possibility of intravenous transplantation of HUCBMSCs in the treatment of cerebral edema secondary to hematoma in neonatal rats. METHODS: Neonatal rats aged 7 days were randomly assigned into three groups: normal control, model group and MSCs transplantation group. One week after transplantation, the rats in each group were randomly killed to extract brain tissues which were used pathomorphology observation and index measurement. RESULTS AND CONCLUSION: Both brain coefficient and water content of brain tissue were significantly lower in the MSCs group than the model group (P < 0.01). However, there was no difference between MSCs group and control group (P > 0.05). There was no difference in the death rate of rats in each group 1 week after the implantation. The brain damage to most of the rats in this group was not serious but only slight by chance, close to the normal at the one week after implantation. HUCBMSCs transplantation could effectively decrease the brain edema and ameliorate the damaged brain tissue. One week after transplantation, the MSCs appeared in the ischemic damaged brain tissue as a large number of MSCs could cross the blood brain barrier to distribute and scatter around the disease focus integrated with host brain tissue without apparent threshold. Related Articles | Metrics

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Bone marrow mesenchymal stem cells protect against renal ischemia reperfusion injury through paracrine mechanism Hu Hong-lin, Wang Gong-xian, Zou Cong, Huang Xue-ming, Xi Xiao-qing, Shi Zi-min 2011, 15 (32):  5995-5998.  doi: 10.3969/j.issn.1673-8225.2011.32.024 Abstract ( 251 )   PDF (550KB) ( 432 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can be used in the animal experiments of renal ischemia reperfusion injury. OBJECTIVE: To explore the paracrine mechamism of BMSCs in treatment of renal ischemia reperfusion injury. METHODS: Rat BMSCs were isolated by percoll density gradient centrifugation and labeled with 4,6-diamidino-2-pheny-lindole (DAPI). Male SD rats were subjected to clamping renal pedicles for 45 minutes and divided randomly into the transplanted group and the control group. DAPI-labeled MSCs or saline were transplanted into the rats by inferior vena cava. Immunohistochemical method was used to detect the expression of vascular endothelial growth factor (VEGF) and tumor necrosis factor alpha (TNF-α) in kidneys day 2 post-operation. RESULTS AND CONCLUSION: Compared with the control group, the serum creatinine and blood urine nitrogen in the transplanted group were significantly decreased at 1 and 2 days after operation (P < 0.05). At 3 days, the DAPI-positive cells could be found in the renal tissue in the transplanted group and significantly increased at 4 days(P < 0.05). The expression of VEGF was increased and the expression of TNF-α was decreased in kidneys in the transplanted group at 2 days by immunohistochemical method. BMSCs can protect against renal ischemia reperfusion injury through the paracrine mechamism. Related Articles | Metrics

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Bone marrow mononuclear cell transplantation combined with core decompression treatment for early osteonecrosis: A three-dimensional gait analysis Zhao De-wei, Xie Hui, Cui Da-ping 2011, 15 (32):  5999-6002.  doi: 10.3969/j.issn.1673-8225.2011.32.025 Abstract ( 284 )   PDF (586KB) ( 339 )   Save BACKGROUND: Gait analysis provides an objective and accurate quantitative assessment for the orthopedic examination. OBJECTIVE: To observe the gait kinematic characteristics and change rule of patients with early avascular necrosis of the femoral head following treatment of bone marrow mononuclear cell (BMMCs) transplantation combined with core decompression treatment.   METHODS: Three-dimensional motion capture gait analysis was performed in 21 cases of early osteonecrosis undergoing BMMCs transplantation with core decompression. Before and 3, 6, 9, 12 months after treatment, step length, step width, angle of hip motion, kinematic data of hip angle acceleration in the 22 patients walking on the treadmill of 0° and 15°. Gait data of 33 healthy subjects were collected for comparison. RESULTS AND CONCLUSION: Three months after treatment, 21 patients presented with reduced pain gait. The spatial-temporal parameters had no difference from normal people at 3 months for treadmill movement of 0° and 6 months for treadmill movement of 15°. At 9 months, kinematic indexes for 15° inclined treadmill movement were consistent with the normal. The findings indicated that BMMSCs for necrosis of the femoral head can recover normal gait mode of the hip in a time-dependent manner. Related Articles | Metrics

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Dynamic fluorescence in situ hybridization monitoring for chromosomal changes in patients with leukemia after chemotherapy and stem cell transplantation Zhang Zheng-hao, Qu Jian-hua, Chen-shuang, Liu Yang, Jiang Ming, Duan Xian-lin, Yuan Hai-long, Wang Lei 2011, 15 (32):  6003-6006.  doi: 10.3969/j.issn.1673-8225.2011.32.026 Abstract ( 281 )   PDF (674KB) ( 322 )   Save BACKGROUND: Fluorescence in situ hybridization (FISH) is widely used in the detection of abnormal genes in leukemia, but the application of this technique for comparing the efficacy of chemotherapy and transplantation has not been coverage a lot. OBJECTIVE: To evaluate the monitoring results of FISH in patients with leukemia after chemotherapy and hematopoietic stem cell transplantation. METHODS: A total of 85 specimens from 44 leukemia patients after chemotherapy or allogeneic stem cell transplantation were studied by FISH and conventional cytogenetic analysis. We divided the results into four groups: pretreatment group, chemotherapy group, special drug group (all-trans retinoic acid and imatinib mesylate) and hematopoietic stem cell transplantation group (transplantation group). RESULTS AND CONCLUSION: FISH in the diagnosis and monitoring of leukemia minimal residual diseases was more accurate than conventional cytogenetic analysis; The result of FISH showed that the percentage of positive cells in the special drug and transplantation groups was lower than that in the pretreatment and chemotherapy groups (P  < 0.01), but there was no difference between the special drug and transplantation groups (P  > 0.05). The incidence of acute graft-versus-host disease had no correlation with the results of FISH at 1 month after transplantation (P  > 0.05). The FISH results at 1 month, 1-6 months, over 6 months after transplantation had no difference in statistics (P > 0.05). FISH can detect of chromosomal changes for patients after transplantation accurately and effectively, and it can be used to evaluate the trends of disease progression in patients. Related Articles | Metrics

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Effects of bone marrow mesenchymal stem cells transplantation on angiogenesis and hepatocyte growth factor expression in the rat brain after focal cerebral ischemia Li Guo-qian, Wang Jie-hua, Yang Xiao-xia, Hong Zhu-quan 2011, 15 (32):  6007-6011.  doi: 10.3969/j.issn.1673-8225.2011.32.027 Abstract ( 259 )   PDF (1353KB) ( 395 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) transplantation can improve neurological function in cerebral ischemic rats. However, few studies have reported this mechanism. OBJECTIVE: To probe into the mechanisms of BMSCs transplantation on the recovery of neurological functions in the rat brain after focal cerebral ischemia. METHODS: Male adult Sprague-Dawley rats were randomly assigned into 4 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group, vehicle group and MCAO+BMSCs group. Permanent focal cerebral ischemia models were established using the modified Longa’s method. BMSCs were injected into the lateral ventricle of MCAO+BMSCs group one day after right MCAO, and the same dose of PBS was given to the vehicle group. RESULTS AND CONCLUSION: A lot of microvessels proliferated in the injured cortex, reaching peak at 2 weeks. The microvessel density in the injured cortex of rats treated with BMSCs was higher than that of the MCAO group and the vehicle group (P < 0.01). The expression of hepatocyte growth factor (HGF) in brain tissues in the MCAO+BMSCs group was higher than that in the MCAO group and the vehicle group at 4, 7, 14 days after treatment (P < 0.01). These data suggest that BMSCs transplantation can improve neurological function in cerebral ischemic rats through promoting revascularization in the injured region. Related Articles | Metrics

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Effect of olfactory ensheathing cell transplantation on gene expression of Sema3A and its receptor, NP-1, in rats following spinal cord injury Zhu Zhen-dong, Zhang Lin, Zhou Xue 2011, 15 (32):  6012-6015.  doi: 10.3969/j.issn.1673-8225.2011.32.028 Abstract ( 308 )   PDF (1208KB) ( 321 )   Save BACKGROUND: Studies have proved that olfactory ensheathing cells (OECs) can stimulate the repair of spinal cord injury, but the research for mechanism of nerve growth inhibition guidance factor is rarely reported. OBJECTIVE: To investigate the effects of OECs transplantation on the change of mRNA of Sema3A and NP-1. METHODS: A total of 17 female adult healthy Sprague-Dawley rats were randomly divided into 3 groups: OECs transplanted group, DMEM/F12 transplanted group and normal control group. Transplanted groups were anaesthetized and then were acutely transplanted after the left spinal cord hemisection on the level of T11-12 with suspension of OECs or DMEM/F12 culture fluid. RESULTS AND CONCLUSION: Two segments of the spinal cord adjacent to the hemisected segments were dissected 6 weeks later, and reverse transcription polymerase chain reaction (RT-PCR) was used for semi-quantitative analysis of the mRNA of Sema3A and NP-1. There were a large number of OECs alive 6 weeks later. The mRNA quantity of Sema3A and NP-1 in the DMEM/F12 transplanted group was more than that in the normal control group (P < 0.05). But the mRNA quantity of Sema3A and NP-1 in the OECs transplanted group was markedly down-regulated compared with the DMEM/F12 transplanted group, and there was no significant difference between OECs transplanted group and normal control group (P > 0.05). These indicated that the mRNA expression of Sema3A and NP-1 was effectively decreased by OECs transplantation. Related Articles | Metrics

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Effects of serum containing Bing’s giant salamander on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells Xie Xin-wen, Xu Wei, Li Ning 2011, 15 (32):  6016-6020.  doi: 10.3969/j.issn.1673-8225.2011.32.029 Abstract ( 271 )   PDF (1562KB) ( 509 )   Save BACKGROUND: Bing’s giant salamander has the effect to enhance the fracture healing, but the action mechanism and active constituent are still unclear. OBJECTIVE: To investigate the effects of the serum of Bing’s giant salamander on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs) by different concentrations. METHODS: Rats were given Bing’s giant salamander by high, moderate and low concentration, respectively, and their serum was obtained after 7 days. The control serum was obtained by giving equal volume physiological saline. rBMSCs were obtained from Wistar rats and screened by the adhesive method. The serum was supplemented into the culture medium of third passage rBMSCs by different concentration (2.5%, 5%, 7.5%, 10%). The proliferation of rBMSCs was analyzed by MTT reduction assay. The osteogenic differentiation markers including alkaline phosphatase (ALP) activity, calcium deposition, Collagen Ⅰ, osteocalcin and mineralized bone modulus were compared among the exposed groups and the control. RESULTS AND CONCLUSION: The values of A570 in serum groups containing 5% and 7.5% Bing’s giant salamander were significantly higher than those of other concentrations and the control. 5% was stronger than 2.5% in significantly enhancing the osteogenic differentiation of rBMSCs, indicated by significantly improved ALP activity, calcium deposition, Collagen Ⅰ, osteocalcin and the number of mineralized bone nodules compared to the control and other groups (P < 0.01). The metabolites of Bing’s giant salamander after oral administration stimulate the proliferation and osteogenic differentiation of rBMSCs, indicating that they are effective to treat the patients with delayed fracture healing. Related Articles | Metrics

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Effects of assemble flavone of rhizoma drynariae on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells Liu Wei, Zhao Jin-min, Su Wei, Li Xiao-feng, Qin Yi-wu 2011, 15 (32):  6021-6026.  doi: 10.3969/j.issn.1673-8225.2011.32.030 Abstract ( 292 )   PDF (1916KB) ( 464 )   Save BACKGROUND: Rhizoma drynariae is commonly used in orthopedics and showing an accurate effect; however, it specific mechanisms are not very clear. OBJECTIVE: To explore the influences of assemble flavone of rhizoma drynariae on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs). METHODS: The rabbit BMSCs were isolated from femur and tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferated and purified in vitro. The influence on the rabbit BMSCs proliferation caused by different concentration of assemble flavone of rhizoma drynariae was observed with methyl thiazdyl tetrazolium (MTT) assay; rabbit BMSCs were treated with assemble flavone of rhizoma drynariae inductor in vitro, then to observe the morphology by scanning electron microscope (SEM), identified by alkaline phosphatase staining, and the calcium node formation was detected by alizarin bordeaux staining and von Kossa staining. RESULTS AND CONCLUSION: Rabbit BMSCs could be isolated and cultured by combination of gradient centrifugation and different adherent method. After co-culturing by assemble flavone of rhizoma in vitro, the results of MTT showed that the assemble flavone of rhizoma drynariae with 10-6 mmol/L had an obvious promotion on the rabbit BMSCs proliferation. After assemble flavone of rhizoma drynariae induction, osteoblast-like cell morphology and calcium node formation could be observed under SEM; after dyeing, cells were positive to alkaline phosphatase staining, alizarin bordeaux staining and von Kossa staining. Assemble flavone of rhizoma drynariaecan can accelerate the proliferation and differentiation of rabbit BMSCs. Related Articles | Metrics

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Effects of high glucose and Astragalus Membranaceus on the number, proliferation, and differentiation of endothelial progenitor cells from human peripheral blood in vitro Xu Han-song, Lei Min-xiang, Kong De-ming, Xiang Hui, Xie Xiao-yun 2011, 15 (32):  6027-6031.  doi: 10.3969/j.issn.1673-8225.2011.32.031 Abstract ( 274 )   PDF (1314KB) ( 367 )   Save BACKGROUND: The quantity and impaired function of endothelial progenitor cells (EPCs) is the important link of diabetic vascular complications. Astragalus Membranaceus (AMI) can protect endothelium of dysfunction which caused by various reasons and can effectively protect the function of vascular endothelium. OBJECTIVE: To investigate the effects of high glucose and AMI on the number, proliferation, and differentiation of EPCs from human peripheral blood in vitro. METHODS: EPCs were incubated with various concentrations of glucose for 24 hours and with 30 mmol/L glucose for different periods. Simultaneously, EPCs were pre-incubated with 20 g/L AMI for 24 hours and then all the cells were incubated with 30 mmol/L glucose for 24 hours. RESULTS AND CONCLUSION: High glucose decreased the number of EPCs and abilities of EPCs proliferation and differentiation in a dose- and time-dependent manner (P < 0.05 or 0.01). Pre-incubated with AMI could increase EPCs quantity and improve EPCs proliferative and differentiative capacity (P < 0.05 or 0.01). AMI can increase the number and improve the capacity of proliferation and differentiation of EPCs cultured with high glucose. Related Articles | Metrics

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Effects of Ginsenoside Rg1 on the expressions of Nanog, c-Myc, Oct, Klf4, Sox2 mRNA during the differentiation of bone marrow mesenchymal stem cells into pluripotent stem cells Li Jia-wei, Zhang Mi-xia, Zhou Tao, Torao Ishida, Wang Xiu-yun 2011, 15 (32):  6032-6035.  doi: 10.3969/j.issn.1673-8225.2011.32.032 Abstract ( 294 )   PDF (945KB) ( 439 )   Save BACKGROUND: Somatic cells are reprogrammed to induced pluripotent stem cells (iPSCs), mainly through anti-retroviral program to transfer Oct-4, Sox2, c-Myc, Klf4 and other genes into somatic cells. OBJECTIVE: To observe the effects of Ginsenoside Rg1 on Oct4, Sox2, c-Myc, Klf4, Nanog mRNA expression during differentiation of bone marrow mesenchymal stem cells (BMSCs). METHODS: The BMSCs were cultured; 6.25 μmol/L and 12.5μmol/L Ginsenoside Rg1 of Chinese medicine extraction were added into culture medium, respectively. Oct4, Sox2, c-Myc, Klf4, Nanog mRNA expressions were detected in BMSCs. RESULTS AND CONCLUSION: In the BMSCs cultured by 6.25 μmol/L Ginsenoside Rg1 for 30 days, the expressions of Nanog, c-Myc, Oct, Klf4, Sox2 mRNA of BMSCs were increased, and the expression of the gene Nanog and c-Myc had significantly difference compared with the control group. It demonstrated that Ginsenoside Rg1 could enhance the expressions of Nanog, c-Myc, which are crucial genes on converting the BMSCs to iPS. But Nanog positive iPS cells are very difficult to be discriminated from the embryonic stem cells in gene expression profile; therefore, we consider that Ginsenoside Rg1 might facilitate the differentiation from BMSCs to iPS. Related Articles | Metrics

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Construction and identification of lentiviral vector encoding shRNA against REST/NRSF Li Hong-tu, Shi Ping, Pang Xi-ning 2011, 15 (32):  6036-6040.  doi: 10.3969/j.issn.1673-8225.2011.32.033 Abstract ( 306 )   PDF (1647KB) ( 397 )   Save BACKGROUND: In recent years, many studies showed that respressor element1(RE-1) 1-silencing transcription factor /neurons restrictive silence factor (REST/NRSF) could negative control neurons and islet cell differentiation related gene expression. OBJECTIVE: To construct a lentiviral vector expressing shREST/NRSF. METHODS: Four groups of shRNA sequences specifically targeting the REST/NRSF gene were designed and synthesized and cloned into the pFU-GW-RNAi vector. The obtained lentiviral vector containing shREST/NRSF was confirmed by PCR and sequencing. 293T cells were cotransfected with lentiviral vector L-shREST/NRSF, pHelper1.0 and pHelper2.0.The titer of virus was tested according to the expression level of GFP. And preliminary observations on the situation of transfected rat bone marrow mesenchymal stem cells. Real time PCR was employed to assess the gene silencing efficacy of these recombinants. RESULTS AND CONCLUSION: PCR and DNA sequencing demonstrated that the constructed lentivirus vector L-shNRSE/RE-1 produced REST/NRSF shRNA. And it could be stably transfected to rat bone marrow mesenchymal stem cells, there was the infection efficiency of almost 100%. All of these four shRNAs could achieve gene knock down effect, and 3# shRNA had the most significant gene silence effect among them. The lentivirus vector of shREST/NRSF is constructed successfully. Related Articles | Metrics

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Effects of AMLl-ETO fusion gene on the expression of C/EBPα Zhuang Wen-yue, Chen Zi-xing, Zhao Yun, Cen Jian-nong, Shen Hong-jie, Qi Xiao-fei, Li Bing-zong 2011, 15 (32):  6041-6044.  doi: 10.3969/j.issn.1673-8225.2011.32.034 Abstract ( 303 )   PDF (474KB) ( 378 )   Save BACKGROUND: Recent studies have shown that function of C/EBPα gene can be inhibited by AML1-ETO resulting in differentiation block. OBJECTIVE: To investigate the effects of AML1-ETO fusion gene on transcription factor C/EBPα which is crucial for the differentiation of granulocytes, and to explore its role in leukemogenesis. METHODS: The expression of cell differentiation antigen including CD11b and CD14 was detected by flow cytometry. C/EBPα gene expression was assessed by quantitative PCR, and C/EBPα protein expression was tested by Western blot. The chromatin immunoprecipitation (ChIP)-based PCR was used to investigate the direct interaction between the AML1-ETO and C/EBPα promoter in AML1-ETO positive leukemia cell line. RESULTS AND CONCLUSION: The expression of cell differentiation antigen CD11b and CD14 significantly decreased in transfected cells. C/EBPα expression in both mRNA and protein levels decreased in U937 cells transfected with pcDNA3.1-AML1-ETO. The enriched regions in transfected cells were located within C/EBPα promoter. Results indicated that C/EBPα is the direct target gene of AML1-ETO. AML1-ETO can down-regulate the expression of C/EBPα. Related Articles | Metrics

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Transgenic neural stem cell transplantation for Parkinson’s disease: Possibility and feasibility Ding Ji-gu 2011, 15 (32):  6047-6050.  doi: 10.3969/j.issn.1673-8225.2011.32.036 Abstract ( 319 )   PDF (573KB) ( 441 )   Save BACKGROUND: Which type of nerve cells neural stem cells differentiate into is not only decided by cell own gene regulation, but also influenced by the external signals. OBJECTIVE: To review the biology characteristics, differentiation and transgenic transplantation of neural stem cells in treatment of Parkinson’s disease. METHODS: A computer search of PubMed (1992-01/2009-12) and CNKI (2003-01/2010-12) for articles related to the biology characteristics, differentiation and transgenic transplantation of neural stem cells in treatment of Parkinson’s disease was performed by the fist author using the keywords of “neural stem cells (NSCs), neural stem cell transplant, Parkinson's Disease (PD)” in English and Chinese, respectively. Finally, 30 articles were included. RESULTS AND CONCLUSION: Treatment of Parkinson’s disease with transgenic transplantation of neural stem cells is the most promising treatment option. Under the low oxygen condition, neural stem cells receiving transgenic culture can be induced highly to differentiate into dopaminergic neurons that provide a cell source for treatment of Parkinson’s disease. However, the biological characteristics and induced differentiation of neural stem cells needs further studies. Simultaneously, the biological safety during human body treatment gradually attracts more attentions, such as oncogenicity that is studied further. Related Articles | Metrics

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Advances of stem cells transplantation for spinal cord injury Liu Chang-lu, Wu Yan 2011, 15 (32):  6051-6055.  doi: 10.3969/j.issn.1673-8225.2011.32.037 Abstract ( 284 )   PDF (812KB) ( 679 )   Save BACKGROUND: Stem cell transplantation as the most promising treatment for spinal cord injury, has been confirmed in a large number of animal experiments and clinical trials. OBJECTIVE: To review the related research progress of stem cell transplantation for spinal cord injury. METHORDS: A computer-based online search of related articles were performed in CNKI and Medline database published from January 2006 to December 2010, with the key words of “Stem cells; Spinal cord injury; Cells transplantation” in Chinese and English, respectively. 494 articles were collected, and 24 were included. RESULTS AND CONCLUSION: Studies show that transplanted cells can migrate, differentiate into neurons and secrete neural nutrients to promote nerve tissue repair and improve neural function. Embryonic stem cells have been used in the treatment of spinal cord injury, but the potential tumorigenicity limits its clinical application. Neural stem cells are theoretically preferred treatment of spinal cord injury, but this technique develops slowly because of strict separation and purification technology and expensive cost. Mesenchymal stem cell has rich sources, and simple sampling, with feasible autologous transplantation, which avoids the ethical problems and graft rejection. Currently mesenchymal stem cell has been identified as an ideal autologous stem cell transplantation source. With advancing cell transplantation, gene modification and tissue engineered scaffold transplantation for treatment of spinal cord injury, the application range and treatment effect of Schwann cells and olfactory ensheathing cells are also improved. Related Articles | Metrics

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Biological characteristics and clinical application of skin-derived precursor cells Li Zong-zhe, Li Yang, Zong Zhao-wen, Zhang Lian-yang 2011, 15 (32):  6056-6059.  doi: 10.3969/j.issn.1673-8225.2011.32.038 Abstract ( 324 )   PDF (571KB) ( 385 )   Save BACKGROUND: Skin-derived precursor cells (SKPs) are a novel population of neural crest-related precursor cells that can be isolated from embryonic and adult skin, and have multiple differential potential. OBJECTIVE: To review the cultivation, separation, biological characteristics and application prospects of SKPs. METHODS: The PubMed database was searched for articles published from January 2001 to January 2011. The keywords were “skin-derived precursor cells, skin-derived precursors, SKPs”. The language of articles was limited to English. RESULTS AND CONCLUSION: SKPs are rich in source, easily available and multi-potent. SKPs can differentiate into Schwann cells, neuronal cells and even islet-like cells that have in vitro and in vivo function. SKPs have a good prospect as an experimental and therapeutic resource for disease modelling and regenerative medicine. However, there are still many problems in application. Related Articles | Metrics

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Stem cell therapy for critical limb ischemia Wu Feng-jie, Wang Qiao-yun 2011, 15 (32):  6060-6063.  doi: 10.3969/j.issn.1673-8225.2011.32.039 Abstract ( 317 )   PDF (752KB) ( 458 )   Save BACKGROUND: Recently, stem cell therapy focuses on augmenting neovascularisation and ameliorating collateraI circulation. And it will be a new limbsalving option for critical limb ischemia patients. However, there are few reviews on research progress and development direction. OBJECTIVE: To present a survey of recent publication concerning about stem cell therapy in patients with critical limb ischemia. METHODS: PubMed database and CNKI database were retrieved for publications of critical limb ischemia and stem cell therapy published from 2010-10 to 2011-01. The keywords were “limb ischemia, stem cells transplantation, neovascularisation” in Chinese and “peripheral arterial disease, stem cells transplantation, neovascularzation” in English. Repetitive studies were excluded. A total of 68 literatures were primarily obtained, while 24 were selected for summarization according to inclusion criteria. RESULTS AND CONCLUSION: Critical limb ischemia is the end stage of peripheral arterial disease, with a deep impact on patient's quality of life. In some patients, there are no revascularizing treatment options. Stem cell therapy focusing on augmenting neovascularisation and ameliorating collateraI circulation has raised much interest in the past decade. It is a new limbsalving option for critical limb ischemia patients. However, the underlying mechanisms of neovascularization are still incompletely understood. Both fundamental research as well as large randomized trials are needed for further optimisation of this treatment option, and will hopefully lead to much more benefits for more critical limb ischemia patients in the near future. Related Articles | Metrics

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Biological characteristics and progression of human umbilical cord mesenchymal stem cells Ma Xi-hui, Feng Kai, Shi Bing-yi 2011, 15 (32):  6064-6067.  doi: 10.3969/j.issn.1673-8225.2011.32.040 Abstract ( 589 )   PDF (603KB) ( 1267 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells are sorts of somatic stem cells with the ability of self-renewal and multi-differentiation. It has advantages such as plentiful source, no effect on the donor, convenience for collection and transportation, no heterogenous rejection, and no ethical dispute. OBJECTIVE: To review in the biological characteristics and progression of the human umbilical cord mesenchymal stem cells. METHODS: CNKI, Wanfang, and PubMed databases were searched by computer with key words of “human umbilical cord mesenchymal stem cells, biocharacteristics, gene analysis, induce differentiation” both in English and Chinese. RESULTS AND CONCLUSION: The shape of human umbilical cord mesenchymal stem cells is like typical fibroblast. The antigens of cell surface are various. The cells high-express interstitial cell mark and integrin receptor, otherwise the cells do not express hematopoietic system mark, allostimulatory molecule CD80, CD86 and CD40, human leukocyte antigens HLA-DR, HLA-G, HLA-DP, HLA-DQ, endothelium mark CD31 or CD33, CD14, CD56 and so on. As hematopoietic stem cells and embryonic stem cells, human umbilical cord mesenchymal stem cells could not only differentiate into osteocytes, cartilage cell, hepatic cells and cardiac muscle cells in vitro, but also differentiate dopaminergic neuron, skeletal muscle cell, endothelial cell, islet cell and so on in vivo. But the research of human umbilical cord mesenchymal stem cells has some problems that need to be resolved extremely. For example, the normalization of separation method and culture condition, how to control the growth and differentiation of human umbilical cord mesenchymal stem cells and so on. Related Articles | Metrics

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In vitro induction and differentiation of adipose-derived stem cells Ren Yi-zhong, Han Chang-xu 2011, 15 (32):  6068-6071.  doi: 10.3969/j.issn.1673-8225.2011.32.041 Abstract ( 280 )   PDF (708KB) ( 388 )   Save BACKGROUND: For the past few years, the application of adipose derived stem cells (ADSCs) as seeded cells of tissue engineering is an active field in medications and ADSCs could differentiate into many tissues in vitro. OBJECTIVE: To summarize the situation and progression about ADSCs induced in vitro at home and abroad. METHODS: Databases of CNKI and PubMed (2000-01/2010-10) were selected to search the related articles about ADSCs using the keywords of “adipose-derived stem cells, cell differentiation in vitro, seeded cell, gene transfaction” in Chinese and English. There were 112 articles after the initial survey. Then 28 articles related to sustained-release antimicrobials were involved. RESULTS AND CONCLUSION: ADSCs have extensive self-renewal capacity and the ability to differentiate along multiple lineages. With the multilineage differentiation potential, ADSCs can differentiate not only into such mesenchymal lineages as adipogenesis, chondrogenesis, osteogenesis, myogenesis, but also into neurogenesis of origin ectoderm. In addition, ADSCs have high infected efficiency by oncoretroviral vectors and have the ability to support hematopoiesis. Related Articles | Metrics

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Cardiac electrophysiological characteristics after transplantation of differentiated bone marrow mesenchymal stem cells Liu Bo-wu, Lü An-lin, Yan Xue-bo, Huang Wei, Hou Jing, Li Yao 2011, 15 (32):  6072-6076.  doi: 10.3969/j.issn.1673-8225.2011.32.042 Abstract ( 228 )   PDF (307KB) ( 421 )   Save BACKGROUND: With the development of biotechnology, the electrophysiology of repairing heart tissues of myocardial infarction or myocardial hypertrophy by using bone marrow mesenchymal stem cell has become a hot spot. OBJECTIVE: To overview the research progress of cardiac electrophysiological characteristics after transplantation of induced differentiation of bone marrow mesenchymal stem cells into cardiomyocytes. METHODS: The databases of PubMed, Springer Link, Science Direct and CNKI were retrieved for papers published from January 2000 to October 2010 with the key words of “bone marrow mesenchymal stem cells, cardiac/heart, electrophysiology/electrophysiological characteristics”. The relevant articles concerning cardiac electrophysiological characteristics of induce differentiation and transplantation of bone marrow stem cells were collected. RESULTS AND CONCLUSION: Totally 208 papers have been searched. Preliminary screening by reading abstracts to exclude 162 papers that study purpose do not coincident with this review either contents duplicated, and internalized 46 papers at last. Bone marrow mesenchymal stem cells after induced differentiation and transplantation could improve heart function of animal experimental model and myocardial infarction or myocardial hypertrophy patients. Although the cardiomyocyte-like cells from bone marrow mesenchymal stem cells could help to improve heart function, the cardiac electrophysiological characteristics may be influenced by them. Related Articles | Metrics

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Application of seed cells and scaffolds in the construction of tissue-engineered cornea Tan Xiao-chen, Zhu Huang 2011, 15 (32):  6077-6080.  doi: 10.3969/j.issn.1673-8225.2011.32.043 Abstract ( 241 )   PDF (373KB) ( 377 )   Save BACKGROUND: Tissue engineering has a promising prospect in corneal transplantation. Scaffolds are always restricting the development of tissue-engineered cornea. OBJECTIVE: To analyze application of different seed cells and scaffolds, and to summarize the progress of tissue-engineered cornea in recent years. METHODS: First author searched literature from CNKI (2000/2010-10) and PubMed database (2000/2010-10). The key words are “tissue engineering, corneal transplantation” in Chinese or English. A total of 223 literatures were seized by computers, according to the inclusion criteria, papers concerning the advance, application and reconstruction of tissue-engineered cornea were analyzed. Finally, 33 papers were included for further analysis. The present study was to analyze the seed cells, scaffolds, organ building and clinical applications of tissue-engineered cornea and investigate the development direction in the future. RESULTS AND CONCLUSION: The results show that seed cells and scaffolds are the focus of the studies now. Corneal transplantation has a high success rate in organ transplantation since the immune privilege of eye and avascular cornea, and it will be able to be a tissue engineering organ that can be largely built, easy to transplant. Reconstruction of cornea has reached first base, but every kind of scaffold has certain drawbacks, so the next goal is to find an ideal scaffold material. Related Articles | Metrics