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02 April 2011, Volume 15 Issue 14 Previous Issue    Next Issue

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Differentiation of rat bone marrow mesenchymal stem cells into endothelioid cells in myocardial infarction micro-environment in vitro    Sun Jin-jin, Liu Chao-zhong, Zhang Hai-tao, Luo Hui-lan, Tian Jian-wei, Huang Cong-chun 2011, 15 (14):  2471-2476.  doi: 10.3969/j.issn.1673-8225.2011.14.001 Abstract ( 256 )   PDF (1646KB) ( 370 )   Save BACKGROUND: Derived from the mesoderm, bone marrow mesenchymal stem cells (BMSCs) have the ability of differentiation into a variety of tissues. BMSCs can differentiate into endothelial cells under certain conditions to improve regeneration of blood vessel and repair of injured myocardium. OBJECTIVE: On the bases of establishment of BMSCs cultural system, myocardial infarction (MI) model and differentiating system into endothelial, this study was going to investigate the characters of BMSCs differentiating into endothelial cells and to initially observe the influence of infarcted myocardial micro-circumstances on BMSCs differentiation into endotheliod cells in vitro. METHODS: The bone marrow was extracted from the thighbone and tibiae of SD rats. BMSCs was isolated from rat marrow, cultured and expanded in vitro. Through evaluating the change of morphology, ability of multiplication and differentiating abilities into osteoblast and adipocytes, BMSCs were identified. After a rat MI model was made and myocardial tissue extract of different time after MI: 8 hours, 3 days, 1 week, 4 weeks was prepared, these different tissue extract was combined with the endothelial inducing system including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and insulin growth factor-1 (IGF-1) to induce BMSCs. After 2 weeks, the influence of different inducing circumstances on BMSCs expansion and differentiation into endothelium was observed. Special antigen of Von Willarbrand factor (vWF) was tested at the same. Main outcome measures include: (1)The shapes of the cells, growth curves and the abilities of differentiating into osteoblast and adipocytes were observed; (2)Induced cells:①growth curves; ②the expression rate of the antigen vWF through immunohistochemical; ③RT-PCR method used to test vWF-mRNA expression through semi-quantitative analysis. RESULTS AND CONCLUSION: It was confirmed that the cultured cells was BMSCs and could differentiate into osteoblasts, adipocytes and endotheliod cells. Partial BMSCs induced by growth factors showed positive of vWF and infarcted myocardium tissue extract have cooperative effect with these factors that could improve the process of BMSCs differentiating into endothelial cells (P < 0.05). RT-PCR showed BMSCs induced by infarcted myocardial tissue extract of 1 week could improve this differentiation into endothelium had the highest vWF positive rate. BMSCs are another kind of marrow stem cells apart from hematopoietic stem cells and have ability of multi-potential differentiation into many kind of lineage including endothelium. The induce system contained infarcted myocardium tissue extract could improve BMSCs differentiating into endothelial cells in vitro. This ability may cause BMSCs to play an important role in the repair of injured myocardium and may be the major mechanism of improving the cardiac function during cell therapy. The optimal time of cell therapy is probably at 1 week after MI. Related Articles | Metrics

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Effect of dexamethasone on bone marrow mesenchymal stem cells apoptosis in rats Fan Qie, Zhao Jin-min, Su Wei, Li Xiao-feng 2011, 15 (14):  2477-2481.  doi: 10.3969/j.issn.1673-8225.2011.14.002 Abstract ( 343 )   PDF (1236KB) ( 396 )   Save BACKGROUND: Several studies have shown that dexamethasone can selectively promote bone marrow mesenchymal stem cells (BMSCs) apoptosis. OBJECTIVE: To observe the effect of dexamethasone on BMSCs apoptosis and mechanism of action. METHODS: BMSCs (the first generation) were cultured in vitro; BMSCs of passage 2 cultured were divided into 2 groups: control group and experimental group. Dexamethasone was not added into control group, 10-9, 10-8, and 10-7 mol/L concentration of dexamethasone was added into experimental group, respectively. Apoptosis rate was detected by flow cytometer after 3, 6, 12, 24 hours. MTT assay was used to detect the effect of dexamethasone on proliferation rate of BMSCs. Effects of BMSCs which were deal with by 10-7 mol/L concentration of dexamethasone after 24 hours on Caspase-3 and bcl-2 were measured through RT-PCR. RESULTS AND CONCLUSION: Compared with the control group, 10-8, 10-7 mol/L concentration of dexamethasone had significant promotion on the apoptosis of BMSCs after 12-24 hours, which was relative to concentration of dexamethasone and time. With the effect of increasing concentration and prolonged time, the apoptosis rate of BMSCs had an increasing trend. The results of MTT showed that 10-8, 10-7 mol/L concentration of dexamethasone had significant effect on the proliferation rate of BMSCs after 24 hours, which was consistent with the above-mentioned detection results of flow cytometer. RT-PCR indicated that the activity of Caspase-3 was increased significantly and activity of bcl-2 was decreased mildly. The results confirmed that dexamethasone promoted apoptosis of BMSCs through extracellular pathway and (or) mitochondria pathway. Related Articles | Metrics

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P53-P21 protein pathway impacts on proliferation and apoptosis of 5-azacytidine induced bone marrow mesenchymal stem cells Yan Xue-bo, Lü An-lin,Liu Bo-wu, Hou Jing, Huang Wei, Li Yao 2011, 15 (14):  2482-2486.  doi: 10.3969/j.issn.1673-8225.2011.14.003 Abstract ( 245 )   PDF (1411KB) ( 392 )   Save BACKGROUND: 5-azacytidine (5-AZA) is an effective reagent to induce bone marrow mesenchymal stem cells ( BMSCs) differentiating to cardiomyocytes. OBJECTIVE: To investigate PFT-α blocked P53-P21 protein pathway impacts on proliferation and apoptosis of 5-AZA induced BMSCs. METHODS: BMSCs were isolated from bone marrow of SD rats by density gradient centrifugation. The third passage cells were divided into 4 groups: (1) control group (CON); (2) 5-AZA group; (3) PFT-α group; (4) PFT-α + 5-AZA group. Morphological changes of BMSCs were observe, surface antigen of BMSCs with flow cytometry was identified, proliferation capacity of BMSCs in each group was determined by MTT, apoptosis rate of BMSCs in each group after induction was detected with flow cytometry, and P53, P21 protein expression of BMSCs in each group after induction was determined with the Western blotting method. RESULTS AND CONCLUSION: BMSCs of primary culture form colonies at 2 weeks, passaged cells became larger, elongated spindle, arranged the same trend. The results of Flow cytometry showed CD44 expression of BMSCs was (89.98±1.29)%, CD45 positive expression rate was (2.14±0.22)%. MTT results showed that when the concentration of PFT-α was equal to or less than  20 μmol/L, promoted proliferation of BMSCs, and when concentration reached 40 μmol/L, significantly inhibited proliferation of BMSCs; in 5 days, 5-AZA group significantly inhibited proliferation of BMSCs. Compared with the control group, PFT-α group and 5-AZA + PFT-α group, there was a significant difference (P < 0.01); in 7 days, it also showed such differences (P < 0.01), and BMSCs proliferation of PFT-α group and 5-AZA + PFT-α group was significantly better than blank control group (P < 0.01). The results of Flow cytometry showed, BMSCs apoptosis rate of 5-AZA group was the highest as compared with that of the control group, PFT-α group and 5-AZA + PFT-α group (P < 0.01). Western blotting results showed there was P53, P21 protein expression in each group, the expression of 5-AZA group was significantly increased, P53, P21 protein expression of PFT-α group and 5-AZA+PFT-α group was significantly reduced. By P53 inhibitor PFT-α blocking, P53-P21 protein pathway can significantly reduce the apoptosis of 5-AZA-induced BMSCs, and promote proliferation of 5-AZA-induced BMSCs. Related Articles | Metrics

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Expression of Hedgehog signaling molecules in the osteogenesis of rat bone marrow mesenchymal stem cells  Zhang Xue-ping, Ma Xiao-li, Wang Yun-shan, Shen Hong, Jia Yan-fei, Zhao Yun, Zheng Yan, Xiao Dong-jie 2011, 15 (14):  2487-2490.  doi: 10.3969/j.issn.1673-8225.2011.14.004 Abstract ( 241 )   PDF (1201KB) ( 371 )   Save BACKGROUND: Hedgehog signaling pathway is an important signal pathway, which regulates the development of variety of tissues and organs in the embryonic stage. It also plays an important role in osteogenesis. However, there are few data about the role of Hedgehog signaling molecules in osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. OBJECTIVE: To investigate the expression of Hedgehog signaling molecules during osteogenic differentiation in rat BMSCs in vitro. METHODS: BMSCs were derived from rat bone marrow, cultured and induced by dexamethasone. Identification of osteoblasts was performed by immunohistochemistry. Level of Sonic hedgehog (SHH) and Indian hedgehog (IHH) during osteogenic differentiation of BMSCs were detected by Western Blot. RESULTS AND CONCLUSION: BMSCs were successfully obtained. Expression of collagen type I was increased on days 7, 14 and 21 following dexamethasone induction. In the osteogenic differentiation, a higher expression of SHH protein was detected in osteoblast induced group compared with non-induced group at the same time point (P < 0.05). While IHH expression was decreased and significantly lower in osteoblast induced group compared with non-induced group (P < 0.05). The results indicate that Hedgehog signaling molecules involved in the osteogenic differentiation of BMSCs induced by dexamethasone, and SHH and IHH had difference in osteogenic differentiation of BMSCs.  Related Articles | Metrics

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Induction and differentiation of bone marrow mesenchymal stem cells into hepatocyte-like cells Fan Jing-jing, Li Dong-liang, He Xiu-hua, Cheng Bian-qiao, Ma Ming, Zhang Zhi-qiang 2011, 15 (14):  2491-2494.  doi: 10.3969/j.issn.1673-8225.2011.14.005 Abstract ( 266 )   PDF (1052KB) ( 437 )   Save BACKGROUND: The research has demonstrated that hepatocyte and fibroblast growth factor and other factors can induce bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells in vitro, but the optimum environment of induction is still in the exploratory stage. OBJECTIVE: To observe the differentiation ability of BMSCs into hepatocyte-like cells by hepatocyte growth factor (HGF) and fibroblast growth factor 4 (FGF-4) and other factors. METHODS: BMSCs of rats were isolated and proliferated by the method of whole bone marrow attachment culture. At the third passage, BMSCs were differentiated into hepatocyte-like cells by HGF, FGF-4, insulin iron-selenium transducin, and hexadecadrol in vitro. The morphological changes of cells were observed at 7, 14, and 21 days. The mRNA expression of albumin (ALB), alpha-fetoprotein (AFP) and cytokeratin 18 (CK 18) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Glycogen was tested by periodic acid-schiff (PAS) at 14 and 21 days after induction, and the expression of ALB was detected by immunofluorescence method. RESULTS AND CONCLUSION: With the prolonged induction time, BMSCs showed hepatocyte-like, round cells appeared and showed colony growth, liver cell-specific markers gradually emerge and mature. The AFP mRNA expression was appeared at 7 days after induction, and increased significantly at 14 days, then decreased at 21 days. Cells expressed CK18, ALB mRNA and glycogen at 14 days, and increased with the prolonged induction time. The expression of ALB in the cytoplasm is positive at 14 days after induction, and increased significantly at 21 days. BMSCs have the ability of differentiating into hepatocyte-like cells by the method of HGF, FGF-4 and other factors. Related Articles | Metrics

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Interleukin 6 induces the differentiation of bone marrow mesenchymal stem cells into hepatocytes in vitro Song Li-hua, Wang Yong-jin, Wang Jin-sheng, Huang Yan, Wu Cui-ling, Shi Bian-hua 2011, 15 (14):  2495-2499.  doi: 10.3969/j.issn.1673-8225.2011.14.006 Abstract ( 234 )   PDF (1573KB) ( 504 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are an important source of hepatocytes. BMSCs can differentiate into hepatocytes under the induction of multiple cytokines and growth factors, and participate in repair and reconstruction of liver function. OBJECTIVE: To explore the effect of interleukin 6 on the differentiation of BMSCs into hepatocytes. METHODS: BMSCs were isolated from Kunming male mice and cultured by attachment method and treated with interleukin 6 (2.5, 5, 10, 20 µg/L) in HEPATOZYME-SFM nutrient solution to induce the BMSCs differentiation into hepatocytes. On days 0, 7, 14, 21, 28 after induction, the expressions of cytokeratin 18, alpha-fetoprotein and glycogen were identified by immunohistochemical method and periodic acid-schiff. The contents of albumin and nitric oxide in the liquid supernatant were identified by ELISA and nitrate reductase method, respectively. RESULTS AND CONCLUSION: Interleukin 6 resulted in dose-dependent (2.5, 5, 10 µg/L) and time-dependent increase in the expressions of cytokeratin 18 and glycogen and the content of albumin. The expression of alpha-fetoprotein increased then decreased to disappearance on day 21. The above effects of interleukin 6 (20 µg/L) was weaker than that of interleukin 6 (10 µg/L). The content of small amount nitric oxide was detected when interleukin 6 was 10 µg/L on day 28. These demonstrated that interleukin 6 is able to induce the differentiation of BMSCs into hepatocyte-like cells, which shows strongest effects at 10 µg/L. Increased mass concentration showed decreased induction effects. Related Articles | Metrics

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Effect of human acellular amniotic membrane matrix on synapsin expression of umbilical cord blood mesenchymal stem cells Hu Wei, Yang Feng, Tang You-jia, Chen Xi-lan, Yang Bo, Guan Fang-xia 2011, 15 (14):  2500-2503.  doi: 10.3969/j.issn.1673-8225.2011.14.007 Abstract ( 212 )   PDF (1151KB) ( 320 )   Save BACKGROUND: The effect of natural extracellular matrix on neural development of adult stem cell is a hot research spot currently. More studies on neural markers expression at home and abroad, few studies on synapse development. OBJECTIVE: To investigate the effect of human acellular amniotic membrane matrix on synaptophysin expression of umbilical cord blood mesenchymal stem cells. METHODS: The 3rd generation of human cord blood mesenchymal stem cells in experimental group was seeded in 24 orifice plate of coating membrane-like matrix cover glass. Time was counted when the inoculation, cells were cultured by basal medium for 4 days. There was no membrane-like matrix, the remaining procedures was the same as the experimental group. Morphologic changes of cells were observed by hematoxylin-eosin staining. Synaptophysin-related protein expression (SYN expression) was detected by immunohistochemical SP method at 2 and 4 days inoculation. SYN expression Ⅰ, Ⅱ, Ⅲ of genetic transcription level was detected by RT-PCR method at 2 and 4 days before and after inoculation. RESULTS AND CONCLUSION: Cells in control group was fibroblast-like, cells in experimental group extended the processes and connected each other. SYN expression in experimental group was higher compared with control group; there were significant differences (P < 0.01). SYN Ⅰ, SYN Ⅱ, SYN Ⅲ did not express in transcriptional level both before inoculation and at 2, and    4 days in control group. At 2 and 4 days in experimental group, SYN Ⅱ had low expression, SYN Ⅲ had poor expression, and SYN Ⅰ did not express in transcriptional level. The results indicated that acellular amniotic membrane matrix can promote the production and extension of the umbilical cord blood mesenchymal stem cells process, and connected each other, express SYN, and provide favorable conditions for neural differentiation of umbilical cord blood mesenchymal stem cells. Related Articles | Metrics

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Isolation, culture and biological characteristics of umbilical cord blood mesenchymal stem cells  Sun Guo-jun, Zhao Wen-jing, Chen Xi, Ye Dong-xia, Li Jing 2011, 15 (14):  2504-2507.  doi: 10.3969/j.issn.1673-8225.2011.14.008 Abstract ( 316 )   PDF (1262KB) ( 488 )   Save BACKGROUND: Studies have found that cord blood contains a large member of hematopoietic stem cells and abundant mesenchymal stem cells. OBJECTIVE: To establish an isolating, culturing and proliferating method for human umbilical cord blood mesenchymal stem cells (HUMSCs) and then to observe cell biological characteristics in vitro. METHODS: HUMSCs were isolated and purified by density gradient centrifugation and adhering to the culture plastic. After successive subculture and amplification, the morphology was observed under microscope and the growth curve was drawn. Cell cycle was determined by flow cytometry; The expression of CD29, CD44 and CD34 in HUMSCs were detected by immunohistochemistry (SP); HUMSCs were induced to differentiate into adipocytes in DMEM with 20% horse serum and identified by oil red O staining; HUMSCs were induced to differentiate into osteoblasts by 0.1 μmol/L dexamethasone, 10 mmol/L β-glycerol phosphate sodium ascorbate and 50 μmol/L ascorbic acid and identified by alkaline phosphatase staining.   RESULTS AND CONCLUSION: High purity HUMSCs were isolated from umbilical cord blood. HUMSCs were spindle cells and their living behavior was quite stable. In vitro subcultured nine generations had no morphological changes and no signs of aging; Passage 3 of HUMSCs could differentiate into adipocytes and osteoblasts; The results showed that density gradient centrifugation, adherent culture and digestion time control can obtain an access to high purity HUMSCs with strong proliferation and passage ability in vitro. Related Articles | Metrics

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Autologous transplantation of bone-marrow mononuclear cells for treatment of diabetic lower limb ischemia in 83 cases  Wang Guang-yu, Zhu Lü-yun, Hou Rong-rong, Ma Li-cheng, Hu Li-ye, Li Xiao-ling, Yang Shao-ling, Shan Wei 2011, 15 (14):  2508-2511.  doi: 10.3969/j.issn.1673-8225.2011.14.009 Abstract ( 267 )   PDF (1377KB) ( 496 )   Save BACKGROUND: Autologous transplantation of bone-marrow mononuclear cells for treatment of diabetic peripheral angiopathy has been widely developed at present, which showed good curative effect on ischemic lesions. Case reports also showed that it could improve symptoms of peripheral neuropathy. OBJECTIVE: To observe the effect of before and after autologous transplantation of bone-marrow mononuclear cells on ischemic lesions and neuropathy index. METHODS: A total of 83 patients (153 legs) with lower extremity ischemia were treated with autologous transplantation of bone-marrow mononuclear cells. 250-400 mL autologous bone marrow blood of each patient was distilled. Mononuclear cells were extracted by the density gradient method and the number of mononuclear cells were 1.01×108-4.96×109 (average 2.04±0.53×108). Mononuclear cells were injected into both lower extremities and foot intramuscular (1-3 mL/site) for 3cm distance after dilution. Ischemic index and neuropathy index were detected at 3, 6 months after transplantation. RESULTS AND CONCLUSION: After autologous transplantation of bone-marrow mononuclear cells, the lower limb blood flow and collateral vessels were increased, walking distance was extended, skin temperature was elevated, and the healing of ulcer was promoted. There were significant differences between pre-transplantation and post- transplantation (3 and 6 months after). After treatment for 6 months, the ational symptom of diabetic peripheral nerve was improved, the conduction velocity of motor nerves and sensory nerves was elevated, and there were significant differences between pre- transplantation and post- transplantation (after 6 months). Related Articles | Metrics

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Isolation, culture, identification and in vivo marker of inguinal adipose mesenchymal stem cells in rats Bi Xiao-juan, Ma Yan, Jiang Ming 2011, 15 (14):  2512-2516.  doi: 10.3969/j.issn.1673-8225.2011.14.010 Abstract ( 429 )   PDF (1393KB) ( 542 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) are easily collected and abundantly cultured, ADSCs can proliferate rapidly when being cultured in vitro, with multi-directional differentiation potential, ADSCs are expected as seed cells for tissue engineering and cell therapy. OBJECTIVE: To culture rat ADSCs in vitro, and to mark and identify its differentiation. METHODS: CollagenaseⅠdigestion was used to isolate ADSCs from rat groin fat pads under sterile condition. ADSCs were passaged and amplified by the trypsin digestion. At the third passage, growth curve of the third passage ADMSCs was drawn by cytometry. Cellular surface antigens HCAM, CD106, CD29, CD49D and CD34 were examined using flow cytometry. Giemsa staining, Feridex-labeled and induced to differentiate into osteoblasts and lipocytes. Oil red staining was used to examine lipocyte induction, alizarin red staining wwas used to examine osteoblast induction. RESULTS AND CONCLUSION: In vitro cultured ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. The third passage ADSCs were positive for CD29, but negative for CD34, HCAM, CD49d and CD106. Curve of growth detection had logarithmic phase and platform phase, the positive expression of Feridex reached 80%. ADMSCs were differentiated into lipocytes and osteoblasts under certain conditions. ADSCs isolated from rat epididymal fat pads can be easily cultured, passaged and amplified, and which can mark alive and differentiate into osteoblasts and adipocytes under certain conditions. Related Articles | Metrics

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Effects of prostaglandin E1 on in vitro proliferation of human adipose derived stromal cells  Yang Huan-na, Cui Zheng-jun, Seung-Kyu Han, Hyup-Woo Lee, Zou Shi-bo, Han Zhao-feng, Zhou Jian 2011, 15 (14):  2517-2520.  doi: 10.3969/j.issn.1673-8225.2011.14.011 Abstract ( 327 )   PDF (1408KB) ( 455 )   Save BACKGROUND: Recently, research and application of human adipose derived stromal cells (ADSCs) has become a hotspot; however, there are few reports addressing prostaglandin E1 (PGE1) effects on ADSCs proliferation. OBJECTIVE: To explore the effects of PGE1 on ADSCs proliferation. METHODS: ADSCs were obtained from human normal adipose tissue after isolation and culture. The cultured ADSCs were treated with five different concentration of PGE1, which were 3×10-6, 6×10-6, 9×10-6, 12×10-6 mol/L, and 0×10-6 mol/L (control group). After 3 days, the cell morphology and proliferation were observed. RESULTS AND CONCLUSION: There was no obervious different among the experimental groups. And the cell number of each experimental group was obervious elevated, compared with that of the control group (P < 0.05). These findings indicated that ① PGE1 promoted the proliferation of ADSC; ② The effect of PGE1 had no dose dependent. Related Articles | Metrics

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Induced differentiation of rat adipose stem cells into nerve cells in vitro Gao Hua, Wang Yu-ling, Yao Ya-ni, Yang Xin-ling 2011, 15 (14):  2521-2524.  doi: 10.3969/j.issn.1673-8225.2011.14.012 Abstract ( 352 )   PDF (1103KB) ( 320 )   Save BACKGROUND: Adipose tissue-derived stem cells (ADSCs) have multiple differentiation potentials, which can differentiate into chondrocytes, osteocytes, adipose cells, and cardiac muscle cells under different induced conditions, in addition, they can also differentiated into nerve cells.  OBJECTIVE: To research the in vitro culture and phenotype of rat ADSCs, and to explore it differentiation potential into nerve cells.  METHODS: Inguinal, epididymis and post peritoneal fat tissues were gotten from rats, ADSCs were isolated and cultivated in media, and their shape was recorded passaged. The third or the forth passage cells were identified by stem cells common marker CD29, CD44 and CD45. ADSCs were induced to nerve cells in a simulated nerve cells habitat with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), and then expressions of the neuron specific markers nestin, NF-200 and GFAP were determined with immunofluorescence. RESULTS AND CONCLUSION: ADSCs were successfully extracted from rat fat tissues, and they could also passage infinitely in a considerable large amount. ADSCs strongly expressed CD29 (99.00±0.12)%, CD44 (95.62±0.68)% and low expressed CD45 (0.13±0.12)%. After neurolin induction without serum, induced ADSCs can express nestin; when have been added serum, most of induced cells expressed NF-200 and GFAP. These indicate that ADSCs have robust abilities of duplication. And it has a bionomics orientable differentiated into nerve cells in neural induction circumstance in vitro. ADSCs may be supplied as a source of cells for nervous system injuries and degenerative disease. Related Articles | Metrics

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Horse serum effects on the differentiation of adipose-derived stem cells into myoblasts Liu Jie, Fu Qiang 2011, 15 (14):  2525-2529.  doi: 10.3969/j.issn.1673-8225.2011.14.013 Abstract ( 330 )   PDF (1343KB) ( 336 )   Save BACKGROUND: 5-azacytidine (5-aza)-induced adipose-derived stem cells (ADSCs) into myoblasts required advanced techniques and high cell viability, and long induction time. If certain additive can shorten induction time, this additive would be significant. OBJECTIVE: To assess the horse serum effects on 5-aza-induced ADSCs into myoblasts. METHODS: The first generation of ADSCs fully covered the 100-mm culture dish was passaged to three 6-well culture dishes. In the experimental group, 5% horse serum + 10 μmol/L 5-aza was added in the culture medium. In the control group, 10 μmol/L of 5-aza was simply added. In the blank group, low glucose DMEM medium alone was used; the necessary conditions of culture, subculture and identification are the same. Morphology was observed daily by light microscopy. Immunofluorescence and flow cytometry were used to test the muscle protein differences on days 7, 14, 28, 35. RESULTS AND CONCLUSION: Immunofluorescence and flow cytometry detection for the expression of corresponding muscle-specific cytoplasmic protein showed that time of inducing into myoblasts in the experimental group was shorter than the control group. Studies primarily suggested that the horse serum can shorten the time in 5-aza inducing ADSCs into myoblasts. Related Articles | Metrics

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Effect of granulocyte-colony stimulating factor on mobilization and recruitment of very small embryonic-like stem cells Guo Hui-juan, Wang Jian, Zhang Xing-xiu, Zheng Min 2011, 15 (14):  2530-2534.  doi: 10.3969/j.issn.1673-8225.2011.14.014 Abstract ( 299 )   PDF (1340KB) ( 599 )   Save BACKGROUND: A population of very small embryonic-like stem cells (VSEL-SCs) is a kind of non-hemopoietic stem cells, which has similar biological characteristics to embryonic stem cells. Currently, most studies concerning VSEL-SCs after myocardial infarction, but mobilization of VSEL-SCs and its effect on repair damaged tissues after acute cerebral infarction remain poorly understood.  OBJECTIVE: To investigate the effects of granulocyte-colony stimulating factor (G-CSF) on mobilization of bone marrow-derived VSEL-SCs, recruitment and mechanism after mouse acute cerebral infarction. METHODS: Mouse middle cerebral artery occlusion (MCAO) model was induced by using the filament occlusion method. G-CSF and normal saline were separately injected to MCAO model. Neurological scales were evaluated. The number of VSEL-SCs mobilized into peripheral blood was checked by fluorescence-activated cell sorting analysis. The level of stromal cell derived factor-1 (SDF-1) in plasma and cerebral tissue were determined by ELISA assay. Positive expression of SDF-1 in ischemic regions was detected by immunohistochemistry. RESULTS AND CONCLUSION: Compared with the normal saline group, the neurological behavioral scale of G-CSF treated group was significantly lower at 108 hours after operation (P  < 0.05). G-CSF groups mobilized more VSEL-SCs into peripheral blood compared with the operated control group at 72 and 108 hours (P  < 0.05). The SDF-1 density significantly increased in the plasma and brain tissues of the G-CSF group than that of the normal saline group (P < 0.05), and positive correlation can be seen between the number of VSEL-SCs mobilized into peripheral blood and the SDF-1 plasma concentration. The positive expression of SDF-1 in the G-CSF group was greater than that of the normal saline group. The results indicated that G-CSF mobilized VSEL-SCs in adult MCAO mouse bone marrow into peripheral blood. The mechanism of beneficial effects of G-CSF possibly related to increase of SDF-1 expression in the infracted tissues and recruitment of more VSEL-SCs through CXCR4/SDF-1 axis. Related Articles | Metrics

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A natural activity factor promotes cardiogenic differentiation of bone marrow derived mesenchymal stem cells Peng Peng, Mo Ye-hua, Lin Xiao-li, Li Zhong-yu, Cheng Lei, Zhou Zhen, Li Ming 2011, 15 (14):  2535-2539.  doi: 10.3969/j.issn.1673-8225.2011.14.015 Abstract ( 316 )   PDF (2345KB) ( 548 )   Save BACKGROUND: Previous studies have demonstrated that the methanol extract of Geum japonicum (EGJ) can effectively promote the cardiogenic differentiation of the exogenous mesenchymal stem cells (MSCs), leading to substantial repair of infarcted heart with cardiac function significantly improved. In this study, the isolation of the active compound, which can induce cardiogenic differentiation of MSCs in cell culture system, was described through a bioassay guided fractionation strategy. OBJECTIVE: To isolate active compound (cardiogenin) from Geum japonicum and to observe its activity in promoting cardiogenic differentiation of MSCs. METHODS: The MSCs were isolated from the bone marrow of SD rats. The isolated MSCs were cultured and treated with EGJ (100 mg/L) and cardiogenin (10 mg/L) respectively. Immunofluorescent staining with antibodies specific for early cardiac differentiation marker (MEF2) and myosin heavy chain (MHC) was performed at 3 and 7 days after treatment respectively to monitor the process of cardiogenic differentiation of MSCs. RESULTS AND CONCLUSION: It was found that approximately 39% of both EGJ and cardiogenin treated MSCs were positively stained with MEF2 at 3 days after treatment. On day seven of the treatment, more than 70% of both EGJ and cardiogenin treated MSCs were positively stained with MHC specific antibodies. The cardiogenin can significantly induce cardiogenic differentiation of MSCs in vitro as does EGJ, but the dose of cardiogenin required for induction of cardiopgenic differentiation of MSCs is much less but the induction efficiency is little higher than that of EGJ.  Related Articles | Metrics

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Expression and secretion of hepatocyte growth factor on bone marrow mesenchymal stem cells stimulated by tumor necrosis factor alpha Lu Zheng-feng, Jiang Hai-xing, Qin Shan-yu, Xiao Jian, Zhang Jun-hong, Meng Yun-chao 2011, 15 (14):  2540-2544.  doi: 10.3969/j.issn.1673-8225.2011.14.016 Abstract ( 203 )   PDF (1577KB) ( 410 )   Save BACKGROUND: Previous studies demonstrated that tumor necrosis factor-α (TNF-α) can stimulate the immunosuppression function of bone marrow mesenchymal stem cells (BMSCs), and promote hepatocyte growth factor (HGF) expression of BMSCs. OBJECTIVE: To investigate whether BMSCs stimulated by TNF-α is an effective way to promote the expression and secretion of HGF in rat BMSCs. METHODS: BMSCs from SD rats were isolated, cultured and purified by the whole bone marrow adherence method, generated to passage3-4 to use. TNF-α pre-stimulated BMSCs, the medium was discarded after 5 hours, and then cultured with fresh medium as experimental group, normal cultured BMSCs as blank group. The morphology of BMSCs was observed by the inverted phase contrast microscope. The expression of surface makers CD 29, CD 34, CD 44 and CD 45 of BMSCs were identified by flow cytometer. HGF mRNA and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. HGF protein level in cell supernatants was determined by ELISA. RESULTS AND CONCLUSION: BMSCs were isolated by the whole bone marrow adherence method with uniform and showed spindle shaped, typical swirl morphology. BMSCs expressed CD29+ (99.45%), CD34- (97.91%), CD44+ (99.52%), CD45- (98.42%); the expression level of HGF mRNA and protein and HGF in cell supernatants increased with time-dependent. The expression levels of HGF mRNA and protein and HGF in cell supernatants in experimental group were significantly higher than that in blank group (P < 0.01). BMSCs stimulated by TNF-α can effectively improve the expression and secretion of HGF. Related Articles | Metrics

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Effects of lateral ventricle transplantation of bone marrow stromal cells on the movement and cognitive function of cerebral ischemic reperfusion rats Gu Ping,Zhang Zhong-xia, Zhang Bao-hua, Wang Yan-yong, Zhang Zhen-qing, Cui Dong-sheng, Geng Yuan, Wang Ming-wei 2011, 15 (14):  2545-2550.  doi: 10.3969/j.issn.1673-8225.2011.14.017 Abstract ( 263 )   PDF (1877KB) ( 383 )   Save BACKGROUND: Some results of improved motor function have been achieved by the transplantation of bone marrow stromal cells (BMSCs) into lateral cerebral ventricle to treat middle cerebral artery obstruction (MCAO) rats, while the cognitive function has not been reported. OBJECTIVE: To observe the movement, cognitive function, infarction size of the MCAO rats, and the migration way of the transplanted BMSCs.     METHODS: MCAO 2 hours reperfusion cerebral ischemia models were manufactured and were randomly divided into 3 groups: BMSCs group, phosphate buffered solution (PBS) group, and model group. The suspension of BMSCs (5 μL, 1.0x106/μL) labled with BrdU and the same volume of phosphate buffered solution were injected into the infarction side lateral cerebral ventricle of BMSCs group and PBS group respectively, while model group and normal group received no treatment. The ability of motor coordination of the MCAO rats was observed by balancing beam experiment, the swimming speed and the ability of space learning and memory were detected by morris water maze test. The infarction size was detected by hematoxylin-eosin staining, and the migration way of the BrdU positive cells was detected by immunohistochemistry staining. RESULTS AND CONCLUSION: The score of the balance beam test in BMSCs group was significantly improved on the 3rd, 7th, 14th day after transplantation (P  < 0.05). On the 7th-10th days after transplantation, the swimming speed in the BMSCs group was significantly faster than that of PBS group (P < 0.05). The escape lantency in BMSCs group was significantly decreased (P  < 0.05), while the percentage of swimming time (T1%), swimming distance (D1%) in the plantform quadrant and the crossing times in BMSCs group were significantly increased (P < 0.05). Typical pathomorph of cerebral infarction could be seen on 7th and 14th day after transplantation in hematoxylin-eosin staining of model group rats. There was no obvious difference among the percentage of infarction area. BrdU staining showed that on the 1st day after transplantation, the positive cells were gathered in the transplanted lateral cerebral ventricle, resided in the wall of the cerebral ventricle in the style of tigh disposed cell clump. On the 3rd day, the most positive cells passed through the cerebral ventricle parietes directily migrated to the ischemic zone in the form of single cell. On the 14th day, the most positive cells were found in the striatum and cortex of the ischemia hemisphere. Results indicated that BMSCs lateral ventricle transplantation 24 hours after MCAO operation can significantly improve the motor coordination ability and the function of space learning and memory of MCAO rats. The infarction size of MCAO rats was not diminished, while the BMSCs with lateral ventricle transplantation can survive and migrate to the striatum and cortical area of cerebral ischemia zone directily. Related Articles | Metrics

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Treatment of traumatic brain injury in rats with intravenous transplantation of adipose-derived stem cells Zhang Tian-xiang, Yang Bo, Guan Fang-xia, Hu Xiang, Du Ying, Qiao Xiao-jun, Tian Yi, Ba Yun-tao, Duan Xiao-bing, Deng Xiao-hui, Gu Chen-xi, Zheng Wen-di 2011, 15 (14):  2551-2556.  doi: 10.3969/j.issn.1673-8225.2011.14.018 Abstract ( 282 )   PDF (1723KB) ( 362 )   Save BACKGROUND: Animal experiments and clinical researches have confirmed that stem cells have a therapeutic effect on the traumatic brain injury. OBJECTIVE: To explore the behaviors of rats of acute traumatic brain injury and expressions of glial fibrillary acidic protein (GFAP), neuronspecific enolase (NSE), and nidogen in brain tissue for the treatment of adipose-derived stem cells (ADSCs) transplantation. METHODS: Brain injury model of SD rats were established by free-fall impact method of epidural, and were randomly divided into 3 groups: control group, transplantation group, and saline group. Brdu-labled ADSCs underwent caudal vein injection at 48 hours brain injury in transplantation group; any treatment was not given for the control group; saline underwent caudal vein injection at 48 hours brain injury in saline group. Nerological severiyt score (NSS) method was applied to evaluate the recovery of neurological function in rats. Migration of ADSCs in brain injury tissue was observed by inverted microscope. The expressions of GFAP, NSE, and nidogen in damaged brain tissue of rats were detected by using of immunohistochemistry method. RESULTS AND CONCLUSION: NSS score in transplantation was significantly higher than that in control and saline groups (P< 0.05). Brdu-labled ADSCs could be found in damaged brain tissue. The expression of GFAP and NSE in transplantation group was significantly lower than that in control and saline groups in damaged brain tissue of rats (P < 0.05), and the rate of decay was rapid. The expression of nidogen in transplantation was significantly higher than that in control and saline groups (P< 0.05). It was confirmed that ADSCs through the blood-brain barrier into the damaged brain tissue to protect neuron, and promote the function of neural development and regeneration in terms of up-regulation of nidogen gene expression. Related Articles | Metrics

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Monosialoganglioside combined with bone marrow mesenchymal stem cells transplantation for the treatment of cerebral infarction in rats Zhang Ting-yong 2011, 15 (14):  2557-2561.  doi: 10.3969/j.issn.1673-8225.2011.14.019 Abstract ( 278 )   PDF (570KB) ( 580 )   Save BACKGROUND: The ganglioside is the only world-recognized repair nerve damage, remodeling sovereign remedy of neural network. Monosialoganglioside (GM1) is one kind of ganglioside. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cells (BMSCs) transplantation combined with GM1 on the treatment of cerebral infarction in rats. METHODS: The middle cerebral artery occlusion (MCAQ) models in Wistar rats were established by suture method, which were randomly divided into 3 groups: cell transplantation group, infarction group and cell transplantation + GM1 group. The concentration of 1×1010/L BMSCs suspension or the concentration of 1 mL cell culture fluid was injected into MCAQ models through caudal vein, at the same time, GM1 aqueous solution or normal saline was injected into MCAQ models through abdominal cavity, once a day, for 3 days. Longa ethology scores were determined at 24 hours, 3 days after vein transplantation, and 1, 2 weeks after MCAQ. Neurological damage was detected. After 3 days treatment, RT-PCR and Western Blot was used to detect changes of AQP4 mRNA expression and protein synthesis. After 2 weeks, the survival of implanted cells and the repair of tissue were observed by BrdU immunohistochemistry and hematoxylin-eosin staining. RESULTS AND CONCLUSION: The neurological dysfunction scores in cell transplantation group + GM1 group were significantly lower than those in cell transplantation group and infarction group at 1 and 2 weeks after transplantation (P < 0.05). At 3 days after transplantation, AQP4 protein in the surrounding tissue cerebral infarction and its mRNA expression in infarction group were higher than those in cell transplantation and cell transplantation + GM1 groups (P < 0.05). The number of neurons in BrdU immunohistochemistry and hematoxylin-eosin staining slice in cell transplantation + GM1 group was higher than that in cell transplantation and infarction groups (P < 0.05). The results suggested that BMSCs transplantation combined with GM1 can significantly improve neurological function of cerebral infarction in rats. Related Articles | Metrics

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Stereotactic transplantation of bone marrow mesenchymal stem cells for learning and memory ability of vascular rats Li Li-jie, Kong Fan-ming, Zhang Xue-bin, Wang Xin-ping, Wang Shi-min, Tang Fan, Zhang Wen-zhi, Su Xin 2011, 15 (14):  2562-2566.  doi: 10.3969/j.issn.1673-8225.2011.14.020 Abstract ( 274 )   PDF (678KB) ( 459 )   Save BACKGROUND: The key of stem cells for treating nervous tissue injury is the transplantation of stem cells that have regeneration capacity. The damaged neurons are restored by multiple action mechanisms, which recover the nervous function. OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cells (BMSCs) on learning and memory disorders of vascular dementia model rats. METHODS: BMSCs were separated with density gradient centrifugation and screening the cells adherent to the plastic surface. 10 mg/L BrdU was used to label before cell transplantation. Adult female Wistar rats were used to establish vascular dementia rat models by Permanent bilateral carotid artery ligation, and they were then randomly divided into control group, saline group and transplantation group. In the transplantation group, BMSCs were transplanted into the damaged spinal cord by stereotaxis at day 7 following damage. In the saline group, an equal volume of saline was utilized. In the control group, the rats were left intact. In the transplantation group, BMSCs were transplanted into the damaged Hippocampus by stereotaxis at day 14 following damage. In the saline group, an equal volume of saline was utilized. In the control group, the rats were left intact. Spatial learning and memory was detected by the Morris Water maze. Rats were sacrificed at day 90. Brdu+neuron specific enolase, Brdu+glial fibrillary acidic protein (GFAP) immunohistochemistry double-staining cells, and the migration of Doublecortin (DCX, the marker of N Ps) from the subventricular zone to the damaged hippocampus were observed. RESULTS AND CONCLUSION: ①Spatial learning and memory in rats (escape latency and the number of cross platform) was better in the transplantation group than in the control group (P < 0.05). No significant difference was determined in the control group and the saline group. ②Double-staining cells of immunohistochemistry could be found at the center of damage site and the surrounding site in rats of the transplantation group. Results indicated that BMSC transplantation can improve the recovery of nervous function of rats with spinal cord injury. Its mechanism may be correlated with the differentiation of transplanted cells into neuron-like and glial cell-like cells, secretion or promoting secretion of neurotrophic factors in host. Related Articles | Metrics

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Hematopoietic stem cell transplantation and platelet transfusion Song Kui, Guo Zi-wen, He Hui-qing, Jian Li, Qiu Da-fa, Huang Gui-nian, Lin Shu-hua, Ren Zhi-juan, Li Wei-chao, Liu An-li, Xu Xiao-jun 2011, 15 (14):  2567-2571.  doi: 10.3969/j.issn.1673-8225.2011.14.021 Abstract ( 448 )   PDF (659KB) ( 538 )   Save BACKGROUND: Platelet transfusions were shown to reduce mortality from hemorrhage in patients with acute leukemia in the 1950s, and the use of the therapy has steadily grown since then. The procedure has become an essential part of hematopoietic stem cell transplantation (HSCT) treatment. However, there are rare studies about the current transfusion practices. OBJECTIVE: To retrospectively analyze the effects of platelet transfusion in 52 patients undergoing HSCT. METHODS: Prophylactic versus therapeutic platelet transfusion was grouped according to platelet counts and bleeding manifestation. The factors potentially influencing platelet transfusion efficacy after HSCT were evaluated. RESULTS AND CONCLUSION: The effective rates of prophylactic and therapeutic platelet transfusion groups were 63.6% and 55.6%, respectively. The effective rates and mean platelet increment in patients undergoing HSCT were 60.9% and 26.8×109/L. Meanwhile, the efficacy of prophylactic and therapeutic platelet transfusion in patients who received autologous transplants was similar in the group of allogeneic HSCT. Coagulation disorder as a independent factor identified by binary Logistic Regression showed influencing the efficacy of platelet transfusion．The platelet transfusion is an effectively therapeutic method for thrombocytopenic patients undergoing HSCT. The effect of prophylactic platelet transfusion is better than therapeutic platelet transfusion in these patients. The difference was not seen between the groups of autologous and allogeneic transplants. The curative effect of platelet transfusion can be influenced by coagulation disorder. Related Articles | Metrics

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Stereotactic transplantation of bone marrow mesenchymal stem cells combined with mild hypothermia for traumatic brain injury Yuan Guo-fu, Li Hong-xing, Yuan Xiong-zhou 2011, 15 (14):  2572-2576.  doi: 10.3969/j.issn.1673-8225.2011.14.022 Abstract ( 309 )   PDF (1513KB) ( 465 )   Save BACKGROUND: In recent years, mild hypothermia (33-35 ℃) in the treatment of brain and spinal cord injury obtains more and more attention. Many clinical studies have shown that mild hypothermia can reduce secondary brain and spinal cord injury, and play a precise protective role in central nervous injury in rats. OBJECTIVE: To determine whether mild hypothermia can enhance curative effects of stereotactic transplantation of bone marrow mesenchymal stem cells (BMSCs) on traumatic brain injury (TBI) in rats. METHODS: BMSCs were cultured by the suspension culture in vitro. The models of severe TBI were made in rats by given 253.31-303.98 kPa power of hydraulic impact. BMSCs were stereotactically transplantated into the brain in the rats with TBI 24 hours after TBI, and the rats were divided into three groups, group A: TBI; group B: BMSCs transplantation group; group C: BMSCs transplantation+hypothermia group. After 3 days, AQP4 expression was determined by Western Blot, the water contents in the brain tissues were measured as wet-to-dry weight ratio. The neurological defect scores were determined after 24 hours, 3 days and 1, 2 weeks after TBI. The immunohistochemistry and pathology were checked after 2 weeks. RESULTS AND CONCLUSION: Three days after brain injury, compared with A group and B group, the expression of AQP4 detected by Western Blot and cerebral edema detected by wet-to-dry weight ratio were decreased (P  < 0.05). The neurological defect scores were significantly lower in group C than those in groups A and B at 1 and 2 weeks after TBI (P < 0.05). The number of neurons in group C was higher than that in groups A and B (P < 0.01). These findings show that stereotactic transplantation of BMSCs combined with mild hypothermia can significantly improve the neurological function in the rats with TBI.  Related Articles | Metrics

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Inhibitory effect of bone marrow mesenchymal stem cells carrying cytosine deaminase gene on glioma cells in vitro Xing Qi, Song Fei, Liu Jian, Ji Guang-chun, Zhang Da-qing, Ma Yu-fang 2011, 15 (14):  2577-2582.  doi: 10.3969/j.issn.1673-8225.2011.14.023 Abstract ( 322 )   PDF (2057KB) ( 569 )   Save BACKGROUND: The bone marrow mesenchymal stem cells (BMSCs) expressing foreign cytosine deaminase gene (gene CD) can effectively transform 5-fluorouracil (5-FC) that was used before chemotherapy into chemotherapeutics 5-FC with cytotoxicity. BMSCs expressing gene CD significantly inhibited the growth of glioma cells in vitro. OBJECTIVE: To explore the effects of BMSCs as gene therapy vector expressing exogenous gene cytosine deaminase gene on the proliferation of glioma C6 cells. METHODS: Mouse MSCs were isolated and cultured. Lentivirus vector combined with cytosine deaminase gene and green fluorescent protein (GFP) was constructed. Using lentivirus packaging, cytosine deaminase gene and GFP were transferred to mouse BMSCs. BMSCs expressing cytosine deaminase gene and GFP were stably obtained and cocultured with glioma C6 cells. Following 5-FC was added to the medium, effects of cytosine deaminase gene on the proliferation of glioma cells were detected using flow cytometry. RESULTS AND CONCLUSION: Lentivirus-mediated cytosine deaminase gene and GFP gene successfully transfected mouse BMSCs and formed C57BL/6 mMSC-codA/eGFP cells. C57BL/6 mMSC-codA/eGFP could induce obvious apoptosis of glioma C6 cells following treatment with 5-FC. The apoptotic rate of C6 glioma cells was 60% (P < 0.05) under the action of 5-FC at a concentration of 1×106 μg/L. It was thus concluded that C57BL/6 mMSC-codA/eGFP could convert 5-FC to 5-FU and had significant restriction, even fatal, influence on the growth of C6 glioma cells. Related Articles | Metrics

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Adipogenic differentiation of bone marrow stromal stem cells intervened with serum containing different concentration Epimedium  Liu Hong-jiang, Liu Hai-quan, Qin Jia-jia 2011, 15 (14):  2583-2586.  doi: 10.3969/j.issn.1673-8225.2011.14.024 Abstract ( 290 )   PDF (1409KB) ( 609 )   Save BACKGROUND: How to inhibit adipogenic differentiation of bone marrow stromal stem cells (BMSCs) and promote the osteogenic differentiation of BMSCs is a new approach to prevention and treatment of osteoporosis. OBJECTIVE: To investigate the effect of serum containing different concentration Epimedium on adipogenic differentiation of BMSCs in rats with osteoporosis. METHODS: Three of 15 SPF-class SD female rats were used to isolate and culture BMSCs. The remaining 12 rats were divided into control group, high dose herb group, medium dose herb group, low dose herb group. Adipogenic differentiation was observed after 7 day’s intervention. RESULTS AND CONCLUSION: Medium and low dose herb groups played the effects on suppressing adipogenic differentiation of BMSCs (P < 0.01, 0.05). There was no significant difference among each herb groups (P > 0.05). The medium dose herb was the best in different concentration dose herb on inhibiting adipogenic differentiation of BMSCs. Related Articles | Metrics

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In vitro culture of smooth muscle progenitor cells from adult human peripheral blood Zhou Xue-kai, Ni Xu-dong, Li Fei, Zhang Rong-qing, Guo Wen-yi 2011, 15 (14):  2587-2591.  doi: 10.3969/j.issn.1673-8225.2011.14.025 Abstract ( 227 )   PDF (1390KB) ( 356 )   Save BACKGROUND: Smooth muscle progenitor cells (SPCs) from human peripheral blood have the capacity to differentiate into smooth muscle cells. OBJECTIVE: To investigate the method of in vitro purifying and culturing SPCs, and to observe differentiation and growth of SPCs. METHODS: Mononuclear cells were isolated from adult human peripheral blood by density gradient centrifugation. After 12 days of culture in vitro, SPCs were identified and sorted by flow cytometry. SPCs were then induced to differentiate and amplify. Inverted microscope was used to observe morphological changes. Smooth muscle cells specific markers (α-SMA, Calponin, SM-MHC) were then checked with immunofluorescent staining and Western blotting. In addition, Cell growth curve of culture was drawn. RESULTS AND CONCLUSION: Four days later, cell colonies were presented. Twelve days later, most adherent cells were showing spindle-shape. CD14/CD105 double positive cells were SPCs, which accounted for (71.8±7.2)% of adherent cells. Twenty-eight days later, cells arranged in order, showing whirlpool-shape. On day 28, immunostaining of adherent cells was positive for α-SMA. Expression of calponin and SM-MHC was found at 14 d and 21 d with Western blotting respectively, and they gradually enhanced. Growth curve of SPCs showed that increased logarithmic phase started on the 6th day, and platform phase started on the 12th days. Results indicated that SPCs could be harvested through culturing mononuclear cells from adult human peripheral blood. They could maintain the ability of proliferation, and differentiate into smooth muscle cells in vitro. So, SPCs could be selected as the seed cells. Related Articles | Metrics

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Isolation and determination of muscle-derived stem cells in the rat penile corpus cavernosum Xu Li-jun, Zhong Long-fei, Shan Yu-xi, Xue Bo-xin, Chen Dong 2011, 15 (14):  2592-2596.  doi: 10.3969/j.issn.1673-8225.2011.14.026 Abstract ( 251 )   PDF (1351KB) ( 353 )   Save BACKGROUND: Previous studies have confirmed that stem cells with stem cell markers and muscle-derived stem cell markers were detected in rat penile corpus cavernosum. OBJECTIVE: To perform isolation and phenotype identification of muscle-derived stem cells in rat penile corpus cavernosum on the basis of previous study. METHODS: The penile corpus cavernosum from 2-month-old Sprague-Dawley rats was digested and dissociated by type Ⅰ collagenase and trypsin. Cells were primarily purified by the Preplate differential adhesion technique. PP1-PP6 adhered cells were obtained and the expressions of stem cell markers were detected by flow cytometry, immunofluorescence and western blotting. RESULTS AND CONCLUSION: The adherent capacity of PP1-PP6 cells was gradually weakened after 6-step differential adhesion. After two to three days, PP6 cells began to be adherent and became round or spindle-shaped. Flow cytometry detected stem cell antigen 1 (+) 5.7%, embryonic antigen (+) 2.6%, and desmin (+) 41.2% in PP6 cells. Immunofluorescence exhibited that only a few cells expressed stem cell antigen 1 and embryonic antigen, showing diffused distribution. Stem cell antigen 1 was mainly expressed in cytoplasm, and embryonic antigen was mainly expressed in nuclei. Cell aggregation expressing desmin showed a large number and mainly expressed in cytoplasm. Very a few double-positive cells, such as stem cell antigen 1/embryonic antigen, stem cell antigen 1/desmin and embryonic antigen/desmin, were detected. No significant expression of stem cell antigen 1 and embryonic antigen protein was determined in PP1-PP5 cells. However, stem cell antigen 1/embryonic antigen protein was detected in PP6 cells. Desmin expression was gradually enhanced in PP1-PP6 cells. Results indicated that the cells from rat penile corpus cavernosum had the cell phenotype of stem cells and muscle-derived stem cells. Related Articles | Metrics

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Differentiation of human dermal-derived multipotent stem cells into epidermal cells in diabetic wound Gao Xin-yu, Zhao Li-ping, Ke Chang-neng, Wang Ming-gang, Zhong Xiao-hong 2011, 15 (14):  2597-2600.  doi: 10.3969/j.issn.1673-8225.2011.14.027 Abstract ( 300 )   PDF (1335KB) ( 528 )   Save BACKGROUND: Differentiation of stem cells is closely related to the microenvironment. Therefore, it is assumed that human dermal-derived multipotent stem cells may have a potential to differentiate into epidermal cells in skin wound. OBJECTIVE: To explore the feasibility of human dermal-derived multipotent stem cells differentiating into epidermal cells under the microenvironment of diabetic wound. METHODS: Human dermal-derived multipotent stem cells were isolated and cultured. Diabetic naked mouse models were prepared. The third to fifth generation of human dermal-derived multipotent stem cells labeled with 5-bromodeoxyuridine (5-BrdU) were injected into the skin tissue surrounding the wound of diabetic mouse models. The specimens were harvested from the wound tissues for paraffin embedding, and serial sections were made at the 2nd and 3rd week after transplantation for BrdU and keratin staining. RESULTS AND CONCLUSION: Not only BrdU positive cells aggregated in the epidermis but also some positive cells expressed keratin simultaneously in the epidermis. During the course of diabetic wound healing, human dermal-derived multipotent stem cells have the potential to differentiate into epidermal cells. Related Articles | Metrics

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A rapid method for harvesting murine bone marrow cells using retrograde isolation of the femur and tibia Fu Bi-mang, Su Ying-zhen, He Xiao-wen, Hu An-bin, He Xiao-shun, Huang Jie-fu 2011, 15 (14):  2601-2604.  doi: 10.3969/j.issn.1673-8225.2011.14.028 Abstract ( 680 )   PDF (1161KB) ( 914 )   Save BACKGROUND: There are no detailed reports about how to rapidly isolate muscle tissues from the femur and tibia which directly influences the ischemic period of bone marrow cells in mice. OBJECTIVE: To investigate the rapidest and feasible method for harvesting murine bone marrow cells (BMCs) in order to decrease the ischemia injury of BMCs. METHODS: Muscle and connective tissues surrounding the femur and tibia were entirely removed by using an initiated retrograde method by which dissection of bones were started at ankle joints and ended at hip joints. The key point, in the whole isolating procedure, was “crossing → slipping → replicating” with appropriate force with ophthalmic scissors worked in coordination with fine tissue forceps. Time of entirely isolating the femur and tibia as well as time of harvesting BMCs were recorded. The obtained BMCs were cultured in 25 cm2 corning flasks using RPMI1640 with 10% fetal bovine serum, and after 72 hours, the cells were stained with trypan blue and the survival rate was detected with cell counter. RESULTS AND CONCLUSION: Time of isolating the femurs and tibia and time of harvesting BMCs were (1.9±0.4) minutes and (14.7±1.8) minutes, respectively. No bone or marrow cavity was injured or contaminated. Compared with before culture, the number of BMCs after 72 hours increased significantly [(8.1±2.0)×107 vs. (4.7±0.7)×107, P < 0.01], while the cell survival rate did not change remarkably [(97.5±1.1)% vs. (95.4±1.0)%, P > 0.05]. No visible microbe was found in the culture media. To harvest mouse BMCs using the retrograde isolation of the femur and tibia is a practical, feasible and rapid method. Related Articles | Metrics

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Comparison of in vitro culture of mesenchymal stem cells in the amniotic fluid from different duration of pregnancy Guo Su-hong, Yuan Chun-hua, Liu Hui-ling, Li Xin, Xu Hui-jing 2011, 15 (14):  2605-2608.  doi: 10.3969/j.issn.1673-8225.2011.14.029 Abstract ( 365 )   PDF (642KB) ( 503 )   Save BACKGROUND: Mid-half amniotic fluid was usually collected for mesenchymal stem cells (MSCs) culture, but the study on the other phase amniotic fluid-MSCs was seldom. The culture effect of amniotic fluid from different duration of pregnancy with different media is not reported. OBJECTIVE: To compare the in vitro culture effect of MSCs in the amniotic fluid from different duration of pregnancy. METHODS: Two media were used to isolate and culture MSCs from 60 cases amniotic fluid of pregnancy 15-42 weeks. The growth state and the success rate of MSCs cultivation course were observed. The cytochemical staining was applied to detect the chemical characteristics of MSCs. Growth curve of MSCs after passage was detected by MTT assay. RESULTS AND CONCLUSION: The success rates of 37-42 weeks pregnancy amniotic fluid MSCs were 8% and 44% using PRMI-1640 and AmnioMAXⅡ complete medium containing 10% fetal bovine serum, that of 21-36 weeks were 20% and 60%; but that of 15-20 weeks was 100% using any medium. The result of cytochemical staining showed that POX and SB were negative; PAS, ACP, AENE were positive; NAP was 1% positive. No difference was determined in staining results at different duration of pregnancy. MTT assay showed that growth curve of MSCs exhibited “S” shape. Amniotic fluid-MSCs of 15-36 week pregnancy were transferred for 15 passages, showing prosperity; that of 37-42 weeks could only be transferred for 10 passages, showing slow growth. The results demonstrated that AmnioMAXⅡ complete medium can elevate obviously the success rate of amniotic fluid-MSCs from late-phase amniotic fluid. The optimal collection time of amniotic fluid was 15-20 weeks. Related Articles | Metrics

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Methylation status in the promoter region of secreted frizzled related protein 5 gene in patients with myelodyplastic syndrome Shi Xiao-hong, Fan Yun, Zhou Chang-hu, Ning Shang-yong, Sun Liang, Zhu Xiao-quan, Tang Lei, Chang Nai-bai, Yang Ze 2011, 15 (14):  2609-2612.  doi: 10.3969/j.issn.1673-8225.2011.14.030 Abstract ( 315 )   PDF (670KB) ( 398 )   Save BACKGROUND: Methylation in the promoter region of secreted frizzled related protein 5 (SFRP5) gene is closely related to acute myeloid leukemia. As a pre-leukemia, methylation status in the promoter region of SFRP5 gene in patients with myelodyplastic syndrome (MDS) remains unclear. OBJECTIVE: To investigate the methylation status in the promoter region of SFRP5 gene in patient with MDS. METHODS: Methylation specific PCR (MSP) method was applied to examine the promoter methylation of SFRP5 gene in 43 bone marrow or peripheral blood samples of MDS patients, and 70 normal peripheral blood samples from volunteers of general outpatients as controls. RESULTS AND CONCLUSION: All 113 samples were involved for the results analysis. In 43 patients of MDS, 5 samples (11.6%) showed SFRP5 gene methylation. In 70 controls, 1 sample showed SFRP5 gene methylation. And all of them were part-methylation status. The frequency of SFRP5 gene methylation in MDS patients was significantly higher than that in controls (χ2=5.511, P=0.019). Of the 43 MDS patients’ samples, 17 cases were bone marrow samples, 26 cases were peripheral blood samples. Of the 5 SFRP5 gene methylation samples, 2 cases were bone marrow samples and 3 cases were peripheral blood samples. There was no significant difference between the different sample source (bone marrow or peripheral blood ) for the results of the methylation status (χ2=0.001, P=0.982). The methylation status of SFRP5 gene was correlated with MDS. The methylation of SFRP5 gene may be one of the molecular mechanisms that contributes to the prognosis of patients with MDS. Related Articles | Metrics

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Induced differentiation and malignant transformation of bone marrow mesenchymal stem cells Yuan Xiao-gang, Lü Nong-hua 2011, 15 (14):  2615-2618.  doi: 10.3969/j.issn.1673-8225.2011.14.032 Abstract ( 270 )   PDF (613KB) ( 496 )   Save BACKGROUND: Previous studies have demonstrated that bone marrow mesenchymal stem cells (BMSCs) can undergo malignant transformation and become the origin of malignant tumor cells. OBJETIVE: To review the research progress in the methods and directions of induced differentiation and malignant transformation of BMSCs in recent years. METHODS: The Pubmed database (http://www.ncbi.nlm.nih.gov/pubmed), SPRINGER database (http://www.springerlink.com) and Elsevier database (http://www.sciencedirect.com) were searched online with the key words of "mesenchymal stem cells" AND "differentiation" or "bone marrow mesenchymal stem cells" AND "transformation" in English. Papers were included from January 2000 to November 2010, Language was limited to English. Following reading titles and abstracts, repetitive researches and Meta analysis were excluded, 31 related articles recently published or in authoritative journals were reviewed. RESULTS AND CONCLUSION: There are 3 approaches can be used for inducing BMSCs to differentiate in vitro at least, such as certain conditioned induction mediums, co-culture with purpose cells and cells or serum derived from pathological damaged model. BMSCs can be directionally differentiationed into almost all human tissue types’ cells that from the endoderm, mesoderm and ectoderm. Experimental researchs and clinical case reports suggest that BMSCs can undergo malignant transformation and become the cell origin of many tumors. It is indicated that the security problems of BMSCs in clinical application need further study, and it may also provide a new vision for future tumor biological research. Related Articles | Metrics

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Senescence mechanism and immortalization of human bone marrow mesenchymal stem cells Huang Wei, Lü An-lin, Liu Bo-wu, Hou Jing, Li Yao, Yan Xue-bo 2011, 15 (14):  2619-2623.  doi: 10.3969/j.issn.1673-8225.2011.14.033 Abstract ( 504 )   PDF (579KB) ( 651 )   Save BACKGROUND: Human bone marrow mesenchymal stem cells (hBMSCs) due to its property are considered one of the most promising stem cell for regeneration of the injured myocardiocytes, and have become the hot point for the study of myocardial tissue engineering. OBJECTIVE: To retrospectively analyze senescence mechanism of hBMSCs and to establish immortalized method of hBMSCs according to these mechanisms. METHODS: The first author retrieved PubMed Database, Wanfang Database, Springer and Science Direct Database for articles on senescence mechanism of hBMSCs and the immortalized method published from 1990 to 2010. RESULTS AND CONCLUSION: The senescence and loss of the stem cell property of BMSCs were found during the cell cultivation in vitro, which profoundly hinders clinical application of hBMSCs. In light of the mechanism of senescence including p53-p21 and p16-pRb, researchers have put intensive effort in finding ways to build up immortalized hBMSCs; methods for immortalization involving KP cells that transfect HPV16 E6E7 gene into BMSCs or 3A6 cells that both HPV16 E6E7 gene and phTERT -IRES2-EGFP are transfected into hBMSCs. Methods above had prolonged the BMSC survival. However, DNA transfection-based methodologies entail some risk of genomic recombination or insertional mutagenesis as well as innate antiviral responses. Thus, the success rate of transfection is low. The search for ways to avoiding senescence without incurring genetic change has become the focus of intense research effort. Standard methods for building up immortalized BMSCs require further study. Related Articles | Metrics

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Inhibitory effects of umbilical cord blood hematopoietic stem cells on primary hepatocellular carcinoma Chen Yi-ming, Li Li, Ran Jiang-hua 2011, 15 (14):  2624-2628.  doi: 10.3969/j.issn.1673-8225.2011.14.034 Abstract ( 318 )   PDF (615KB) ( 321 )   Save BACKGROUND: Primary hepatocellular carcinoma is a common malignant tumor in clinic. Although various treatment measures such as surgery, radio frequency ablation, selective arterial embolization have made progresses, these treatments are not suitable for a few patients who suffered from advanced hepatocellular carcinoma. Under such circumstances, more researchers are concerned with the therapeutical or inhibitory effects of stem cells on malignant tumors. OBJECTIVE: To review incidence, immunoediting process, microenvironment of primary hepatocellular carcinoma and suppressive effect of hematopoietic stem cells on it or other solid malignant tumors. METHODS: Computer-based online search of Wanfang database and Foreign Medical Journal full-text Service database was performed for literatures concerning incidence, immune microenvironment of primary hepatocellular carcinoma and suppressive effect of hematopoietic stem cells on solid malignant tumors including hepatocellular carcinoma published between January 2000 and September 2010. The key words were “hepatocellular carcinoma, hematopoietic stem cell, microenvironment”. Finally, 33 literatures were included. RESULTS AND CONCLUSION: Occurrence and development of the tumor closely associated with changes in the body immune system. In the body with a tumor, the immune system is under inhibitory state, and has no response or low response to malignant tumor cells. Umbilical cord blood hematopoietic stem cells were characterized by low immunogenicity, strong fission ability and wide differentiation lineage. Umbilical cord blood hematopoietic stem cells can differentiate into various lineages of cells in the body following transplantation, including many immune cells, which can supplement the defects of the immune system, and kill tumor cells, and inhibit tumor growth. However, its action mechanism, indication and adverse effect need more explorations. Related Articles | Metrics

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Research progress in tumor stem cells Hou Ping, Li Jian-ping 2011, 15 (14):  2629-2632.  doi: 10.3969/j.issn.1673-8225.2011.14.035 Abstract ( 399 )   PDF (569KB) ( 733 )   Save BACKGROUND: Tumor stem cells have a strong proliferation and differentiation, but different from normal stem cells.These stem cells also has developed into tumor, so they are widely concerned by scholars. OBJECTIVE: To review the characteristic of tumor stem cells and the relationship with stem cells and tumor, and clinical significance. METHODS: A computer-based online search of PubMed database, CNKI database, Wanfang database for articles of tumor stem cells was performed with the key words of “tumor stem cell” in Chinese and English, respectively. A total of 89 literatures were retrieved, and 30 documents of which were included. RESULTS AND CONCLUSION: At present, tumor stem cell is still in the stage of basic research in China, while tumor stem cell has made some progress on experimental research and clinical application at abroad. It generally believed that tumor stem cells are existed in tumor tissue. Tumor stem cells play an important role in development and treatment of tumor, tumor stem cells for the treatment of tumor are also very optimistic about the prospects of clinical application, but the characteristics of tumor stem cells and relation with adult stem cells are still in the preliminary research stage. Related Articles | Metrics

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Strategy study of treatment for cerebral infarction sequelae using neural stem cell transplantation Chen Xiao-yu, Duan Yun-xia, Fang Liang, Zhang Zi-qian, Liu Qing-shan 2011, 15 (14):  2633-2636.  doi: 10.3969/j.issn.1673-8225.2011.14.036 Abstract ( 317 )   PDF (682KB) ( 679 )   Save BACKGROUND: Studies have demonstrated that stem cell transplantation plays a positive role in treatment of cerebral infarction, which also has good outcome in recovery and diminution of infarct size in clinical trials.  OBJECTIVE: By analyzing and summarizing the research which adopts treatment of cerebral infarction sequelae in past ten years, we devote to discuss the strength and weakness of those treatments and find a suitable treatment of cerebral infarction sequelae. METHODS: Application of computer technology to retrieve the articles in wanfang data and PubMed database on spinal cord tissue engineering from 2000 January to 2010 October. Words for retrieve were “cerebral infarction sequelae, neural stem cells”.  Twenty-two literatures were remained for analysis. RESULTS AND CONCLUSION: Neural stem cell transplantation for treating cerebral infarction sequelae are focus on integration, supplementation, or substitution damaged cells by transplanting exogenous neural stem cells, activation or repair injured endogenous neural stem cells. Neuroprotection aims directly at various passageways during ischemic cascade reactions, protects neurons which damaged by ischemia or hypoxia but has viable to reverse ischemic penumbra, decreases reperfusion injury, and reduces infarct size. Applications of physical therapy, functional rehabilitation and ethnic medicine provide new hope for treatment of cerebral infarction sequelae. How to regulate proliferation and differentiation of neural stem cells in vivo and promote neurologic repair is the research focus. Related Articles | Metrics

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Neural stem cells and Parkinson disease Wei Shen 2011, 15 (14):  2637-2640.  doi: 10.3969/j.issn.1673-8225.2011.14.037 Abstract ( 263 )   PDF (537KB) ( 447 )   Save BACKGROUND: Parkinson disease in the elderly has become a common movement disorders, currently its main form of treatment is still in the fields of medicine and surgery, and the role of neural stem cells in functional recovery in Parkinson's disease has been domestic and foreign scholars’ attention. OBJECTIVE: To explore the effect of neural stem cells on Parkinson disease based on the analysis of biological characteristics of neural stem cells and pathological features of Parkinson disease, in order to provide a scientific basis for rehabilitation of Parkinson disease promoted by neural stem cells transplantation. METHODS: A computer based online search of Medline database, CNKI database and VIP database (1994-01/2010-08) was performed with the key words of "Parkinson disease, neural stem cell, rehabilitation" in English and Chinese, respectively. According to the relevant literature of Parkinson disease and neural stem cells, articles published in recent years or in authorized journals were included. Duplicated and outdated articles were excluded. The standard literatures were involved for analysis. RESULTS AND CONCLUSION: Through research we found that neural stem cells in the rehabilitation of patients with Parkinson disease have many advantages, such as their materials can be easily obtained, low immunogenicity and orientable differentiation. Neural stem cells transplantation has been served as a treatment for nerve damage, including Parkinson disease. But if neural stem cells are applied to clinical treatment, it is still need to be constantly researched. The differentiation and mechanisms of functional repair in neural stem cells are unclear. Whether cells can be integrated with injury in vivo after transplantation to establish normal synaptic connections also need to be further studied. Related Articles | Metrics

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Immunological characteristics and transplantation of bone marrow mesenchymal stem cells in treatment of spinal cord injury  Liu Xiao, Xu Tie 2011, 15 (14):  2641-2644.  doi: 10.3969/j.issn.1673-8225.2011.14.038 Abstract ( 231 )   PDF (593KB) ( 392 )   Save BACKGROUND: Previous studies have confirmed that bone marrow mesenchymal stem cells (BMSCs) can be also transplanted into allogeneic and heterologous gene environment, and maintain multi-directional differentiation potential. It may provide a new path to promote nerve repair after spinal cord injuries for its rich source, avoidance of ethical problem and distinct immunological characteristics. OBJECTIVE: To summarize the immunological characteristics, different pathways of transplantation and its possible mechanisms of BMSCs and to provide theoretical evidence for stem cells in treatment of spinal cord injury. METHODS: Related articles on PubMed from January 2000 to September 2010 were searched by the first author with the key words of “mesenchymal stem cells, bone marrow, immunological characteristics, transplantation, spinal cord injury” in English. Inclusion criterion: the articles related to the immunological characteristics of BMSCs and research progress of its treatment. Exclusion criteria: the articles with repetitive research or Meta analysis. 260 relevant articles were selected and 33 of them were included. RESULTS AND CONCLUSION: BMSCs have low immunogenicity and can suppress the immunoreaction after transplantation. Migration through different methods, choosing the right migration path and timing may provide useful protection for the treatment of spinal cord injury. The mechanism may include neuron substitute, neurotrophic factor secretion, homing effect and so on. With the deep research of BMSCs and the mechanism of spinal cord injury, it will have a bright future in the field of clinical application after spinal cord injury. Related Articles | Metrics

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Identification of hepatocytes differentiated from stem cells  Li De-qiang, Wang Ren-hao 2011, 15 (14):  2645-2647.  doi: 10.3969/j.issn.1673-8225.2011.14.039 Abstract ( 267 )   PDF (988KB) ( 513 )   Save BACKGROUND: Various methods can induce stem cells differentiate into hepatocytes, so we need a set of convincing method to test. OBJECTIVE: To summarize the identification advances in the methods for testing stem cells differentiate into hepatocytes.  METHODS: A computer-based search was conducted in Foreign Medical Journal Full-Test Service and Pubmed for English articles about isolation, cultivation and induction of stem cells published between January 1985 and June 2010 with the key words of “stem cells, induction, differentiation, hepatocytes, identification”. Meanwhile, Chinese Journal Full-text Database was undertaken to identify relevant articles published between January 2000 and June 2010. A total of 43 articles were collected. RESULTS AND CONCLUSION: Stem cells are characterized by plasticity. Stimulating factors in vitro and special microenvironment in vivo could induce stem cells differentiate into hepatocyte precursors and mature hepatocytes. Stem cells can provide a rich source of liver cells and provide seeds cells for us in theory, which can be applied in experiments and clinic. However, whether the induced cells are what we expected needs a set of scientific and systematic detection methods, and we can prove it according to a few characteristics of hepatocytes, such as surface makers, synthetical and metabolic effects. Related Articles | Metrics

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Differentiation of adipose tissue-derived stem cells into myocardial cells and their transplantation for treatment of cardiovascular diseases Wang Lin-lin, Dong Wei-ren, Zhu Yan-fei, Zhou Yao 2011, 15 (14):  2648-2652.  doi: 10.3969/j.issn.1673-8225.2011.14.040 Abstract ( 234 )   PDF (407KB) ( 406 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs), a kind of adult stem cells, possess plasticity and can be induced into myocardial cells under certain conditions. Autologous ADSCs transplanted into the infarct area can differentiate into myocardial cells and vascular endothelial cells to construct new vessels and thereby improve cardiac pump function. OBJECTIVE: To study the factors that influence ADSCs differentiation and transplantation and the current clinical and laboratory research progress of ADSCs transplantation for treatment of cardiomyopathy. METHODS: A computer-based retrieval was performed in Medline (between January 1990 and April 2010), PubMed database, the China Biological Medicine Database (CBM) (between January 1990 and April 2010), and China National Knowledge Infrastructure (CNKI) with the keywords adipose-derived stem cells, myocardial cells, cell differentiation, cell transplantation, cardiomyopathy treatment. RESULTS AND CONCLUSION: A total of 30 articles, consisting of 6 reviews and 24 randomized controlled trials, were obtained. At present, there have been uniform methods of ADSCs isolation and culture, and ADSCs can be effectively proliferated in vitro, but there have been no direct methods to identify these stem cells. ADSCs differentiation can be induced both in vitro and in vivo, besides, with a characteristic of early differentiation. ADSCs transplantation is a more conductive therapy for myocardial disease compared with bone marrow stem cells (BMSCs) transplantation. Different ADSCs transplantation methods should be carried out in different types of cardiomyopathy. Stem cell labeling technique can help to dynamically monitor implanted in vivo. Transplantion of autologous ADSCs is a new way to treating cardiomyopathy. However, for successes in clinical practice, the method to inhibit tumor cells-promoting characteristics is needed to ensure long-term safety of the patients receiving ADSCs transplantation. Related Articles | Metrics

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Factors influencing yields of mesenchymal stem cells derived from human umbilical cord blood Zhang Ji-hua, Sun Kang, Wang Yan, Tian Shao-qi, Xia Chang-suo, Zhang Cai-long, Yu Teng-bo 2011, 15 (14):  2653-2656.  doi: 10.3969/j.issn.1673-8225.2011.14.041 Abstract ( 266 )   PDF (377KB) ( 342 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) exist in umbilical cord blood (UCB), currently, there is not a method to in vitro separate, culture and amplificate human UCB-MSCs effectively. OBJECTIVE: To explore factors that influence yields of UCB-MSCs. METHODS: The relationship between the success rate of yielding UCB-MSCs and several factors, such as gestational ages (≥40 weeks, 37 weeks and ≤32 weeks), the number of mononuclear cells (MNCs) in UCB (≥2.5×109/L, <2.5×109/L), the inoculum density of MNCs (1×107, 1×109, 1×1011/L), the concentration of fetal bovine serum (FBS, 5%, 10%, 15%, 20%) in culture medium, and whether the culture flask being coated with FBS or not beforehand, as well as relationships among these factors were investigated. RESULTS AND CONCLUSION: The success rate of yielding UCB-MSCs was up to 58.3%. The success rate decreased as the gestational ages increasing (P < 0.01). The success rate could be enhanced to 76.9% when the MNCs count was more than 2.5×109/L, and there was significant difference when comparing to that of the group (36.4%) with MNCs count less than 2.5×109/L (x2=8.07, P=0.005). There was a negative correlation between the MNCs count and the gestational ages in the specimens with the same volume of UCB (r=-0.95, P < 0.01). In the group with the cell inoculum density of 1×1011/L, the growth and proliferation of primary and subculturing MSCs were better than that of the groups with the cell inoculum density lower than 1×1011/L. The adherence of MSCs in the group with the culture medium containing 5% FBS happened much later than other 3 groups, while the purity of MSCs in this group was much higher. When comparing the passage rate, there was no significant difference among the 4 groups with different concentration of FBS. In the group of culture flask being coated with FBS beforehand, the purity and proliferation ability of MSCs was higher than that in the groups with culture flask not being coated. It is suggested that culture of UCB-MSCs was influenced by several factors. The success rate could be increased by choosing the fetus with relative lower gestational ages, collecting enough volume of UCB, inoculating cells with a higher density, choosing the medium with lower concentration of FBS, and coating the culture flask with FBS beforehand. Related Articles | Metrics

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Effects of high glucose and advanced glycation end-products on osteogenic differentiation of human adipose-derived stromal cells in vitro Li Dong-song, Li Shu-qiang, Cai Bo, Wang Ping, Feng Wei, Liu Jian-guo 2011, 15 (14):  2657-2660.  doi: 10.3969/j.issn.1673-8225.2011.14.042 Abstract ( 275 )   PDF (550KB) ( 530 )   Save BACKGROUND: Bone metabolism disorder happens in diabetic environment, bone defects in which are difficult to repair. Study addressing osteogenic property of adipose-derived stroma cells (ADSCs) in diabetic environment provides theoretical basis for its application in certain environment. OBJECTIVE: To explore the effects of high glucose (HG) and advanced glycation end-products (AGEs) on osteogenic capacity of human ADSCs. METHODS: 100 mg/L AGEs and 27.5 mmol/L HG were used to simulate in vitro diabetic environment and intervened ADSCs osteogenic differentiation. The cells were divided into 4 groups, with 6 samples in each group. The expression of type Ⅰ collagen was examined by fluorescent immunofluorescence at 21 days after osteogenic induction. The number of calcification nodes was counted under contrast phase microscopy at 14, 21 and 28 days.  RESULTS AND CONCLUSION: Fluorescent quantitation scan showed that the type Ⅰ collagen amount of the AGEs+HG treated group was 2.76 times lower than that of the control group. AGEs+HG reduced the number of ADSCs calcification nodes compared with the control, HG, and AGEs groups, the differences were statistical significant (P < 0.01). AGEs and HG exposure inhibit the cognate osteogenic differentiation of ADSCs, which suggest that AGEs and HG are unfavorable factors that reduce ADSCs osteogenic ability.  Related Articles | Metrics