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02 April 2013, Volume 17 Issue 14 Previous Issue    Next Issue

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In vivo MRI-traced bone marrow mesenchymal stem cell transplantation in myocardial infarction rats Hua Ping, Wang You-yu, Yang Song-ran, Liu Jia-liang, Liu Li-bao, Tao Jun, Yang Yan-qi 2013, 17 (14):  2471-2479.  doi: 10.3969/j.issn.2095-4344.2013.14.001 Abstract ( 385 )   PDF (2864KB) ( 741 )   Save BACKGROUND: The continuous monitoring of the in vivo survival, distribution, migration, proliferation and differentiation of the stem cells is important to evaluate the efficacy and safety of stem cells transplantation.  OBJECTIVE: To observe the distribution and migration of superparamagnetic iron oxide labeled bone marrow mesenchymal stem cells in ischemic myocardial tissue traced with MRI. METHODS: Rat bone marrow mesenchymal stem cells were separated and cultured by direct attachment method. The surface antigens of the bone marrow mesenchymal stem cells were identified. The bone marrow mesenchymal stem cells were labeled by the novel superparamagnetic iron oxide. The feasibility of labeled bone marrow mesenchymal stem cells was determined by MRI in vitro. Trypan blue exclusion test and methylthiazolyldphenyl-tetrazolium bromide colorimetric test were performed to detect the activity and proliferation of the labeled cells. A total of 60 Sprague Dawley rats were divided into three groups, and the rats were used to establish the myocardial infarction model. At 2 weeks after modeling, the rats were re-implanted with phosphate-buffered solution containing labeled bone marrow mesenchymal stem cells, phosphate-buffered solution containing unlabeled bone marrow mesenchymal stem cells and phosphate-buffered solution in the same dose, respectively. All rats in each group underwent MRI at 1 day and 3 weeks after transplantation to dynamically monitor the distribution and migration of transplanted cells. CD90 immunohistochemistry was done according to MRI. RESULTS AND CONCLUSION: After superparamagnetic iron oxide labeling, the Prussian blue staining showed that the blue iron particles were inside the cytoplasm and the labeling rate was 99%. There was no statistically difference of trypan blue exclusion rate and methylthiazolyldphenyl-tetrazolium bromide absorption value between labeled and unlabeled cells. Labeled cells could be detected as low signal intensity on T2WI and T2W/FFE with in vitro MRI. Labeled bone marrow mesenchymal stem cells showed round low signal intensity on T2WI, T2W/FFE at the edge of the infarcted myocardium at 1 day after transplantation, and the initial low signal became obscure gradually, extended and contrast reduced at 3 weeks after transplantation. CD90 immunohistochemistry staining confirmed that transplanted bone marrow mesenchymal stem cells could migrate from the border to the infarcted region. It was feasible to label the bone marrow mesenchymal stem cells with the novel superparamagnetic iron oxide in rats and the labeled bone marrow mesenchymal stem cells could be tracked on MRI in vitro. Figures and Tables | References | Related Articles | Metrics

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Silencing caspase-3 gene effects on the proliferation and apoptosis of rat bone marrow mesenchymal stem cells Liu Jia-liang, Hua Ping, Yang Song-ran, Tao Jun, Jiang Hui-qi, Wang Meng, Yang Yan-qi 2013, 17 (14):  2480-2487.  doi: 10.3969/j.issn.2095-4344.2013.14.002 Abstract ( 416 )   PDF (1667KB) ( 915 )   Save BACKGROUND: Ischemia and hypoxia myocardial microenvironment leads to poor survival of transplanted cells. OBJECTIVE: To investigate the effects of silencing caspase-3 gene on proliferation and apoptosis of rat bone marrow mesenchymal stem cells under ischemia and hypoxia in vitro. METHODS: Lentiviral short hairpin RNA interference vector targeting caspase-3 was constructed and transfected into mesenchymal stem cells as the transgene group. The normal cell group and the empty vector group were as controls. MTS assay was applied to examine the proliferation of cells in each group. The ischemia and hypoxia model was established. The expressions of caspase-3 mRNA and protein were detected by real-time PCR and immunohistochemistry. The apoptotic rates of the cells at different time points (0, 6, 12, 24 and 48 hours) in each group were evaluated by flow cytometry. RESULTS AND CONCLUSION: Recombinant lentivirus was transfected into mesenchymal stem cells successfully and the proliferation activity of the cells was increased (P < 0.05). Compared with control groups, the levels of caspase-3 mRNA and protein in the transgene group were decreased (P < 0.05) under ischemia and hypoxia. Silencing caspase-3 could reduce the apoptotic rate of mesenchymal stem cells (P < 0.05), and the apoptotic rate was increased slowly as ischemia and hypoxia time prolonging. Silencing caspase-3 can increase the growth speed and the anti-apoptosis ability of mesenchymal stem cells under ischemia and hypoxia in vitro. Figures and Tables | References | Related Articles | Metrics

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Adenovirus-mediated human bone morphogenetic protein 2 gene transfects bone marrow mesenchymal stem cells Yin Cheng-hui, Qiu Jun-qin, Zeng Zhao-xun, Chen Zong-xiong 2013, 17 (14):  2488-2494.  doi: 10.3969/j.issn.2095-4344.2013.14.003 Abstract ( 364 )   PDF (1973KB) ( 818 )   Save BACKGROUND: Bone marrow mesenchymal stem cells as the seed cells for repair of bone and cartilage trauma and degeneration have been paid increasing attention. OBJECTIVE: To investigative the effects of human bone morphogenetic protein 2 gene transfection on Sprague-Dawley rat bone marrow mesenchymal stem cells. METHODS: Sprague-Dawley rat bone marrow mesenchyal stem cells were in vitro isolated, purified and amplified. Adenovirus-mediated human bone morphogenetic protein 2 was transfected into bone marrow mesenchymal stem cells. CD90 and CD45 expression levels were tested by flow cytometry. The successfully packaged virus was transfected into bone marrow mesenchymal stem cells and expression of human bone morphogenetic protein 2 gene was confirmed by enhanced green fluorescent protein expression under the fluorescence microscope. Enzyme linked immunosorbent assay was performed to monitor the expression levels of human bone morphogenetic protein 2 and alkaline phosphatase in mesenchymal stem cells. The effect of human bone morphogenetic protein 2 on the proliferation of bone marrow mesenchymal stem cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were successfully harvested from bone marrow of sprague-dawley rats and identified by flow cytometry. After primary culture for 7-10 days, cultured cells displayed typical fusiform shape and the growth status was like “cobblestones” or “whirlpool” under light microscope and could be differentiated into osteoblast-, adipocyte- and neuron-like cells in vitro. After transfection by human bone morphogenetic protein 2 gene, bone marrow mesenchymal stem cells expressed human bone morphogenetic protein 2 and alkaline phosphatase. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that after transfection by human bone morphogenetic protein 2 gene, bone marrow mesenchymal stem cells exhibited a stronger proliferation capacity (P < 0.05). These findings suggest that human bone morphogenetic protein 2 gene-transfected bone marrow mesenchymal stem cells can successively express high level of human bone morphogenetic protein 2 and alkaline phosphatase, indicating that human bone morphogenetic protein 2 gene can significantly promote the proliferation of bone marrow mesenchymal stem cells. Figures and Tables | References | Related Articles | Metrics

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Biocompatibility of an acellular brain tissue scaffold with bone marrow mesenchymal stem cells Ding Ming-xiang, Lin Shao-hua, Li Liang-ming, Hou Bo, Liang Chao-feng, Gong Jin, 2013, 17 (14):  2495-2500.  doi: 10.3969/j.issn.2095-4344.2013.14.004 Abstract ( 428 )   PDF (1695KB) ( 638 )   Save BACKGROUND: Use of scaffolds loaded with bone marrow mesenchymal stem cells for treatment of central nervous system injury is still in the stage of laboratory study. OBJECTIVE: To evaluate the biocompatibility of acellular brain tissue scaffold with rat bone marrow mesenchymal stem cells in vitro and to testify whether the material could be used as a carrier for central nervous system tissue engineering. METHODS: Bone marrow mesenchymal stem cells were isolated and purified by whole bone marrow method, and the acellular brain tissue scaffold was made by both physical and chemical extraction methods. Bone marrow mesenchymal stem cells transfected by green fluorescent protein were cultured and compounded onto acellular brain tissue scaffold in vitro. The microstructure of the materials and the morphological changes of bone marrow mesenchymal stem cells compounded onto the acellular brain tissue scaffold were observed under inverted phase microscope, scanning electron microscope, and confocal laser scanning microscope． RESULTS AND CONCLUSION: The acellular brain tissue scaffold had the structure of three-dimensional network. The cells grew in an adherent way on the acellular brain tissue scaffold and had normal shape in vitro. The rat acellular brain tissue scaffold has good biocompatibility with bone marrow mesenchymal stem cells and might be used as a biological material for central nervous system tissue engineering. Figures and Tables | References | Related Articles | Metrics

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Constructing tissue-engineered adipose with the combination of gene-transfected human umbilical cord mesenchymal stem cells and silk fibroin scaffold Liu Yi, Tang Jun, Li Shi-long 2013, 17 (14):  2501-2508.  doi: 10.3969/j.issn.2095-4344.2013.14.005 Abstract ( 317 )   PDF (2391KB) ( 623 )   Save BACKGROUND: The method of construction of tissue-engineered adipose is imperfect at present. Although the tissue-engineered adipose can be constructed, the efficacy is not satisfactory. OBJECTIVE: To study the new method of construction of tissue-engineered adipose, in order to observe capacity of human umbilical cord mesenchymal stem cells transferred with recombinant insulin gene lentiviral vector combined with silk fibroin scaffold in the construction of tissue engineering adipose in Wistar rats. METHODS: Human umbilical cord mesenchymal stem cells were separated and cultured, and then transfected with recombinant insulin gene lentiviral vector (transfected group) by the best multiplicity of infection =10. The nontransfected human umbilical cord mesenchymal stem cells were regarded as control group. The human umbilical cord mesenchymal stem cells in the transfected group and the control group were seeded onto the silk fibroin scaffold and implanted into the subcutaneous layer of Wistar rats. At 12 weeks after implantation, the transplants were taken, and then identified with fluorescence in situ hybridization and observed with histomorphology and scanning electron microscopy. RESULTS AND CONCLUSION: Oil red O staining showed the transplants in two groups were positive, suggesting that the transplants were synthesized in adipose tissue, and the number of fat-like cells in the transfected group was significantly higher than that in the control group (P < 0.01). Hematoxylin-eosin staining showed significant angiogenesis appeared in or around the new formed tissue, the structure of which was similar to natural adipose tissue. Silk fibroin scaffold in the transfected group was degraded significantly, the number of new vessels in the transfected group was more than that in the control group, and inflammatory cell infiltration in the transfected group was significantly less than that in the control group. Scanning electron microscopy results showed that fat-like cells in the trasnfected group congregated and the structure was similar to that of the normal adipose tissue; the fat-like cells in the control group scattered in the pore of the scaffold. Insulin gene could obviously promote human umbilical cord mesenchymal stem cells to differentiate into adipose; human umbilical cord mesenchymal stem cells transferred with recombinant human insulin gene lentiviral vector composite with silk fibroin scaffolds can construct tissue-engineered adipose in the Wistar rats, and its structure is similar to natural adipose. Figures and Tables | References | Related Articles | Metrics

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Construction of an adenoviral vector encoding Ad5GM-CSF-IL-2 in human bone marrow mesenchymal stem cells Zhang Yu-ping, Zhang Xuan, Guo Zhi, Tan Xiao-hua 2013, 17 (14):  2509-2516.  doi: 10.3969/j.issn.2095-4344.2013.14.006 Abstract ( 359 )   PDF (872KB) ( 817 )   Save BACKGROUND: A problem needed to be solved is how to deliver activating factors for dendritic cells and T cells into the tumors. Owing to the characteristics of tumor tropism and weak immunogenicity, human bone marrow mensenchymal stem cells are used as a vehicle for transferring the activating factor genes of the dendritic cells and T cells to the tumor, which may be a protocol to solve the problem. OBJECTIVE: To construct a type 5 adenoviral (Ad) vector encoding granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-2 (IL-2) genes, and detect the expression levels and duration of GM-CSF and IL-2 after infection of human bone marrow mesenchymal stem cells, providing experimental evidence for in vivo activation of dendritic cells and T cells. METHODS: GM-CSF and IL-2 cDNAs from the total RNA extracted by human peripheral blood mononuclear cells were cloned by RT-PCR and inserted into the eukaryotic expression vector pcDNA3.1/Myc-His(-)B. GM-CSF and IL-2 cDNAs linked by the internal ribosomal entry sites (IRES) from encephalomyocarditis virus were subcloned into the shuttle vector pDC515 for the construction of pDC515 GM-CSF-IRES-IL-2. By using AdMaxTM adenovirus vector system, the shuttle vector pDC515 GM-CSF-IRES-IL-2 and the backbone vector pBGHfrt△E1,3 FLP were cotransfected into 293 cells, and Ad5 GM-CSF-IL-2 was obtained by FLP recombinase-mediated site-specific recombination. The amounts of GM-CSF and IL-2 in the culture supernatants at different time points were measured by enzyme-linked immunosorbent assay after human bone marrow mesenchymal stem cells were infected by Ad5 GM-CSF-IL-2 and irradiated with γ-ray to lose proliferative activity. RESULTS AND CONCLUSION: The sequences of GM-CSF and IL-2 cDNAs were identical with those provided by GenBank NM_000758 (435 bp) and GenBank NM_000586 (462 bp) by sequencing, respectively. AdMaxTM was an efficient and quick packaging system of adenoviral vectors. The GM-CSF and IL-2 linked by IRES in the adenovirus can efficiently be expressed in the human bone marrow mesenchymal stem cells at a high level for 7 days, suggesting a potential application to the tumor immunotherapy taking human bone marrow mesenchymal stem cells as a carrier expressing the activating factors for dendritic cells and T cells. Figures and Tables | References | Related Articles | Metrics

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Recombinant human bone morphogenetic protein-2 adjusts expression of vascular endothelial growth factor in human adipose-derived mesenchymal stem cells Cao Xin, Jin Ge-le, Yang Yi, Chen Hui-jin, Yin Jian 2013, 17 (14):  2517-2524.  doi: 10.3969/j.issn.2095-4344.2013.14.007 Abstract ( 285 )   PDF (860KB) ( 655 )   Save BACKGROUND: Recombinant bone morphogenetic protein-2 can promote tissue engineering bone vascularization, but its biological rules targeting human cells are not clear. At present, there is in which report on recombinant bone morphogenetic protein-2 adjusts the expression of vascular endothelial growth factor in human cells.  OBJECTIVE: To observe and compare the expression of vascular endothelial growth factor in human adipose-derived mesenchymal stem cells on gene level and protein level at different time points after induced with human recombinant bone morphogenetic protein-2.   METHODS: Adipose-derived mesenchymal stem cells were separated from adult human adipose tissues and cultured until passage 3, then divided into induced group and control group. The cells in the induced group were induced by human recombinant bone morphogenetic protein-2 which final concentration was 100 μg/L, then the samples were collected at 3, 6, 12, 18, 24, 36 and 48 hours after induction. Reverse transcription-PCR and enzyme-linked immunosorbent assay were used to detect vascular endothelial growth factor expression on gene level and protein level, compared with the control group. RESULTS AND CONCLUSION: Human recombinant bone morphogenetic protein-2 adjusted vascular endothelial growth factor expression of adipose mesenchymal stem cells in a time-dependent manner, and the expression of vascular endothelial growth factor changed at different time points. Compared with the control group, human recombinant bone morphogenetic protein-2 could suppress vascular endothelial growth factor expression at 3-6 hours (P < 0.05), while at 18-24 hours, human recombinant bone morphogenetic protein-2 could promote vascular endothelial growth factor expression (P < 0.05). These two time periods should be paid attention when using human recombinant bone morphogenetic protein-2 to promote tissue engineering bone vascularization. Figures and Tables | References | Related Articles | Metrics

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Adipose-derived stem cells differentiate into osteoblasts and chondrocytes Liu Xiao-tan, Xu Hai-bin, Lu Tan 2013, 17 (14):  2525-2531.  doi: 0.3969/j.issn.2095-4344.2013.14.008 Abstract ( 446 )   PDF (1772KB) ( 744 )   Save BACKGROUND: Adipose-derived stem cells can be separated and obtained from fat tissue. Fat tissue distributes in the whole body, and can be easily obtained in large quantities and has less damage to the donor site when drawing. OBJECTIVE: To identify the methods of in vitro isolating and culturing of mice adipose-derived stem cells, to induce the adipose-derived stem cells to differentate into chondrocytes and osteoblasts and to investigate the feasibility of being seed cells in tissue engineering. METHODS: The adipose-derived stem cells were isolated from the epididymal fat tissue of Kunming mice. Primary adipose-derived stem cells were obtained and purified by collagenase Ⅰ digestion and differential adherence method. The adipose-derived stem cells were induced with osteogenic induction medium, and then gomori alkaline phosphatase staining and alizarin red calcium nodules staining were performed to detect the differentiation of adipose-derived stem cells; the adipose-derived stem cells were induced with cartilage induction medium, and the toluidine blue staining, safranin-O staining and type Ⅱ collagen immunohistochemistry testing were performed to detect the differentiation of adipose-derived stem cells. RESULTS AND CONCLUSION: The adipose-derived stem cells were spindle-shaped and in adherent growth. After primary cultured for 7-9 days, the cells could reach 90% confluence. After passaged to the third generation, the cell morphology was in consistency, and the growth curve of the passaged adipose-derived stem cells presented “S” shape. The expressions of CD29 and CD44 antigens were positive detected with cell-specific antigen test, but the expressions of CD34 and CD45 were negative. After osteoblast-inducing culture, the differentiation of adipose-derived stem cells towards osteoblasts was verified positively by alkaline phosphatase staining and alizarin red staining. After chondrocyte-inducing culture, the differentiation of adipose-derived stem cells into the chondrocyte was verified positively by oil red O staining, type Ⅱ collagen immunohistochemical staining and toluidine blue staining. The adipose-derived stem cells with differentiation potential can be isolated from the fat tissues, and can be the cells can be stably passaged and differentiated in vitro. The adipose-derived stem cells can be differentiated into osteoblasts and chondrocytes after induction, and the adipose-derived stem cells have the advantages of rich sources and easily obtained which can be considered as the ideal seed cells of tissue engineering. Figures and Tables | References | Related Articles | Metrics

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Signal pathway effects on gastric cancer stem cells and gastric cancer occurrence Chen Hao, Xu Lang 2013, 17 (14):  2532-2537.  doi: 10.3969/j.issn.2095-4344.2013.14.009 Abstract ( 447 )   PDF (1191KB) ( 669 )   Save BACKGROUND: The effect of Hedgehog and Wnt/β-Catenin signal pathway on gastric cancer stem cells and the occurrence and development of gastric cancer is rarely reported. OBJECTIVE: To investigate the mechanism of Hedgehog and Wnt/β-Catenin signal pathway on occurrence of gastric cancer stem cells. METHODS: Gastric cancer stem cells were selected from cell suspension in the culture system of  serum-free medium by magnetic activated cell sorting system. Immunohistochemical SP test was applied to detect the expressions of SHH, GLI1, Wnt2 and β-Catenin protein in gastric cancer stem cells. The relationship among these factors was analyzed by Spearman.   RESULTS AND CONCLUSION: The expression positive rates of SHH, GLI1, Wnt2 and β-Catenin were 74.7%, 78.3%, 85.5% and 83.3% respectively. It was remarkably higher than that of tissues adjacent to cancer (P < 0.05). The expression of each factors had negative relationships (P < 0.05). Hedgehog and Wnt/β-Catenin signal pathway were stimulated abnormally in gastric cancer stem cells. These two pathways may have a co-relationship in gastric cancer occurrence. It provides a new research direction for the treatment of gastric cancer. Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord mesenchymal stem cell transplantation combined with angioplasty for diabetic foot: 3 months angiographic evaluation Qin Han-lin, He Ke-wu, Gao Bin, Ji Ya-li, Huang Yong-cui, Wang Sheng-qin, Hu Guo-ping, Zhou Xue-yong 2013, 17 (14):  2544-2551.  doi: 10.3969/j.issn.2095-4344.2013.14.011 Abstract ( 428 )   PDF (380KB) ( 593 )   Save BACKGROUND: At present, there is no effective drug for the treatment of diabetic arteniosclerosis obliterons, and most of the patients have to receive amputation, and have poor prognosis. OBJECTIVE: To treat the diabetic foot with human umbilical cord mesenchymal stem cells transplantation combined with angioplasty, and to perform the angiographic evaluation at 3 months after operation. METHODS: Forty cases (52 limbs) diagnosed with severe symptoms of Fontain Ⅳ diabetic foot accompanied with varying degrees of lower extremity arterial disease were included. The 12 cases (18 limbs) in the control group received simple interventional treatment, and 28 cases (34 limbs) in the experimental group were given human umbilical cord mesenchymal stem cells endovascular infusion and injection around the ulcer. Then all the patients were followed-up for 3 months.   RESULTS AND CONCLUSION: Local cool-feeling, pain and numbness after treatment were improved continuously in a certain extent; skin temperature, ankle-brachial pressure index and claudication distance were improved persistently. Re-examination angiography at 3 months after treatment showed that the neovessels were increased significantly and the ulcer was healed or decreased gradually. No serious complications and adverse reactions were observed before and after infusion. The results indicate that human umbilical cord mesenchymal stem cell transplantation combined with angioplasty is a safe, effective and satisfactory method for severe diabetic foot. Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cells cultured in artificial meninges repair infarcted myocardium in rats Ma Hong-fen, Zhang Xiao-gang, Shi Ruo-fei, Xiong Ting-lin, Zhao Xia 2013, 17 (14):  2552-2557.  doi: 10.3969/j.issn.2095-4344.2013.14.012 Abstract ( 300 )   PDF (804KB) ( 540 )   Save BACKGROUND: Stem cell transplantation has broad application prospects in the treatment of myocardial infarction. To find the ideal cell type and the effective transplantation method is the key factor to improve. OBJECTIVE: To investigate the effect and safety of artificial meninges combined with bone marrow mesenchymal stem cells on repairing infarcted heart. METHODS: Bone marrow mesenchymal stem cells were obtained by whole bone marrow culture method, and subcultured to the third generation. Then bone marrow mesenchymal stem cells were labeled with 2-(4-amidinophenyl)- 6-indolecarbamidine dihydrochloride and seeded on the cells and artificial meningeal complex. Sprague Dawley rats model of myocardial infarction were established, and 60 rats were randomly divided into sham-operation group, myocardial infarction group, artificial meningeal group, and cells and artificial meningeal complex group. The cardiac function parameters were tested at 4 weeks after transplantation, Western blot was used to test the expression of myocardial tissue connexin 43, and the survival rate was calculated after myocardial infarction. RESULTS AND CONCLUSION: Frozen sections of the heart tissues showed a few blue-stained cells under the fluoroscope in the complex group at 4 weeks after myocardial infarction and transplantation, which indicated that a few bone marrow mesenchymal stem cells survived in the heart. Compared with myocardial infarction group and artificial meningeal group, left ventricular function was improved, the expression of connexin 43 protein and the survival rate were increased in the complex group (P < 0.05). Artificial meninges combined with bone marrow mesenchymal stem cells can improve cardiac function and survival rate in myocardial infarction rats. Figures and Tables | References | Related Articles | Metrics

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Immune tolerance induced by bone marrow mesenchymal stem cells after islet allotransplantation in diabetic minipigs Yang Rui, Ma Zhao-jie, Chen Rong-ping, Wang Mei, Zhang Zhen, Cai De-hong, Chen Hong 2013, 17 (14):  2558-2562.  doi: 10.3969/j.issn.2095-4344.2013.14.013 Abstract ( 264 )   PDF (684KB) ( 433 )   Save BACKGROUND: Islet transplantation is a promising method used to radically cure diabetes mellitus. The immune rejection caused by transplantation hinders the development of transplantation. The immune tolerance of recipients to donor grafts is a key to solving the problem of allotransplantation rejection reaction. OBJECTIVE: To investigate the influence of infusion of bone marrow mesenchymal stem cells on the survival time of islet allograft in the minipigs after islet allotransplantation. METHODS: Under sterile condition, minipig bone marrow was taken and the supernatant was discarded after centrifugation. After washed with L-DMEM culture medium, mononuclear cells were adjusted to be 1×109/L. After 48-hour culture, nonadherent cells were discarded and mesenchymal stem cells were isolated and amplified. Passage 5 cells were taken as donors and cell concentration was adjusted to be 1×1010/L. Minipig models of alloxan-induced diabetes mellitus were prepared and then randomly divided into two groups: bone marrow cells and bone marrow cells + mesenchymal stem cells. In the bone marrow cells group, islet cells and bone marrow cells were infused. In the bone marrow cells + mesenchymal stem cells group, islet cells, bone marrow cells and bone marrow mesenchymal stem cells were infused.  RESULTS AND CONCLUSION: Compared with infusion of simple bone marrow cells, infusion of bone marrow mesenchymal stem cells and bone marrow cells significantly prolonged the survival time of islet allografts (P < 0.05) and maintained high serum insulin level for a longer time in diabetic minipigs. These findings suggest that bone marrow mesenchymal stem cell infusion can significantly prolong the survival time of islet allografts and contribute to the normal function of islet allografts in diabetic minipigs. Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord-derived mesenchymal stem cell transplantation for the treatment of acute lung injury in rats Cui Lei, Lei Xin, Niu Yu-hu, Chen Yan, Niu Bo, Liu Zhuo-la 2013, 17 (14):  2563-2569.  doi: 10.3969/j.issn.2095-4344.2013.14.014 Abstract ( 326 )   PDF (2243KB) ( 457 )   Save BACKGROUND: Mesenchymal stem cells have immunomodulatory properties. Umbilical cord-derived mesenchymal stem cells have unique advantages, and have a bright future in the clinical treatment of acute lung injury and acute respiratory distress syndrome. OBJECTIVE: To investigate the protective effect of umbilical cord-derived mesenchymal stem cell transplantation on endotoxin-induced acute lung injury model. METHODS: Forty-eight healthy male Sprague Dawley rats were randomly divided into normal control group, acute lung injury group and umbilical cord-derived mesenchymal stem cell transplantation group. The rats in the acute lung injury group and umbilical cord-derived mesenchymal stem cell transplantation group were used to establish the acute lung injury model through intratracheal instillation of endotoxin. One hour after modeling, rats in the umbilical cord-derived mesenchymal stem cell transplantation group were instilled with umbilical cord-derived mesenchymal stem cells suspension intratracheally, and the rats in the normal control group and acute lung injury group were instilled with normal saline in the same dose intratracheally. After treated for 24 and 72 hours, the pathological changes of the lung tissue were observed, and the histological score, wet and dry weight ratio of the lung tissue, myeloperoxidase activity and the levels of interleukin-6, interleukin-10 and tumor necrosis factor-α were detected. RESULTS AND CONCLUSION: Umbilical cord-derived mesenchymal stem cell transplantation could attenuate lung injuries in rat models of acute lung injury. Compared with acute lung injury group, the histological score, wet and dry weight ratio of the lung tissue, myeloperoxidase activity and the levels of interleukin-6 and tumor necrosis factor-α in the umbilical cord-derived mesenchymal stem cell transplantation group were decreased at different time points, while the level of interleukin-10 was increased. Umbilical cord-derived mesenchymal stem cells have protective effect on endotoxin-induced acute lung injury models; the protection mechanisms may be that the umbilical cord-derived mesenchymal stem cell transplantation can maintain the balance between inflammatory mediators and anti-inflammatory mediators in lung tissues. Figures and Tables | References | Related Articles | Metrics

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Isolation, culture and identification of rat bone marrow-derived endothelial progenitor cells Wei Xiao-yan, Zhang Li, Lin Ming, Chen Huan, Peng Bo, Peng Yuan-yuan, Li Fu-yun, Hong Pei-xin, Fan Yi-fan 2013, 17 (14):  2570-2577.  doi: 10.3969/j.issn.2095-4344.2013.14.015 Abstract ( 442 )   PDF (2085KB) ( 724 )   Save BACKGROUND: Endothelial progenitor cells have a very broad application prospect, but the isolation and culture methods differ greatly and therefore are hard to repeat. OBJECTIVE: To investigate the isolation and culture methods of bone marrow-derived endothelial progenitor cells. METHODS: Bone marrow mononuclear cells were isolated from 4-week-old Sprague-Dawley rats by density gradient centrifugation methods and cultured by EGM-2 MV medium. Then the cells were identified by morphological observation, FITC-UEA-1 binding and Dil-Ac-LDL uptake assay, and fluorescent immunocytochemistry for detection CD133 and VEGFR2 expression. In addition, angiogenic tube formation was determined by Matrigel tube formation assays. RESULTS AND CONCLUSION: (1) Morphology: After induced culture, the isolated bone marrow mononuclear cells exhibited a spindle-shaped, triangular, and round appearance in the early stage (about the 8th day) and round and short spindle-shaped appearance in the late stage (about the 15th day). (2) FITC-UEA-1 binding and Dil-Ac-LDL uptake assay: Cells were positive on days 8 and 21. (3) Fluorescent immunocytochemistry: on day 8, cells expressed CD133 and VEGFR2. (4) Matrigel tube formation assays: Capillary-like structures formed at 15 hours on Matrigel. These findings suggest that rat bone marrow mononuclear cells isolated by density gradient centrifugation method and cultured by EGM-2MV medium correspond to the characteristics of endothelial progenitor cells. This method is simple, quick, and reliable to harvest enough endothelial progenitor cells. Figures and Tables | References | Related Articles | Metrics

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Endothelial progenitor cells from human umbilical cord blood and peripheral blood: Isolation, culture and identification Li Huan-huan, Zhao Li-jing, He Xu, Wang Juan, Liu Kang-ding 2013, 17 (14):  2578-2585.  doi: 10.3969/j.issn.2095-4344.2013.14.016 Abstract ( 329 )   PDF (521KB) ( 796 )   Save BACKGROUND: There are many articles on the isolation, culture and identification of endothelial progenitor cells at home and abroad, while the articles regarding the endothelial progenitor cells from the umbilical cord blood and peripheral blood are rare. OBJECTIVE: To isolate, culture and identify the endothelial progenitor cells from umbilical cord blood and peripheral blood. METHODS: Mononuclear cells were isolated from umbilical cord blood and peripheral blood with density gradient centrifugation method, and then the cells were implanted on the culture plate pre-paved with fibronectin with the concentration of 1×106/cm2. After that, the cells were induced and cultured with M199 medium containing vascular endothelial growth factor. RESULTS AND CONCLUSION: The endothelial progenitor cells could be isolated and obtained from peripheral blood and umbilical cord blood, and the mononuclear cells obtained with density gradient centrifugation and adherence screening method could be differentiated into endothelial progenitor cells after induced with vascular endothelial growth factor. The endothelial progenitor cells could express CD34, CD133, CD105, KDR and CD13, could swallow acetylated low density lipoprotein and be combined with with ulex europaeus agglutinin 1, which could be considered as the sign of in vitro sorting of endothelial progenitor cells. Figures and Tables | References | Related Articles | Metrics

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Isolation and characterization of amniotic fluid-derived stem cells in Turner’s syndrome Gong Yu, Luo Yu-mei, Tian Lin, Liu Hai-bo, Chen Xin-jie, Sun Xiao-fang, Chen Yao-yong 2013, 17 (14):  2586-2591.  doi: 10.3969/j.issn.2095-4344.2013.14.017 Abstract ( 457 )   PDF (1548KB) ( 474 )   Save BACKGROUND: Great progress has been achieved on studies on isolation, culture and biological characteristics of amniotic fluid-derived mesenchymal stem cells. However, few studies are reported on amniotic fluid-derived mesenchymal stem cells in 45, X/46, XX (Turner’s syndrome). OBJECTIVE: To develop a simple culture protocol to isolate amniotic fluid-derived mesenchymal stem cells and investigate the biological characteristics of amniotic fluid-derived mesenchymal stem cells.   METHODS: We developed a gradient dilution culture protocol to isolate a population of 45, X/46, XX (Turner’s syndrome) amniotic fluid-derived mesenchymal stem cells from second-trimester amniocentesis. The morphology of amniotic fluid-derived mesenchymal stem cells was observed by light microscope. The karyotype was analyzed. Specific cell surface antigens and cell cycle of the clonal amniotic fluid-derived mesenchymal stem cells at passage 4 were characterized by flow cytometry. Osteogenic differentiation of amniotic fluid-derived mesenchymal stem cells was confirmed by alkaline phosphatase staining and alizarin red staining. RESULTS AND CONCLUSION: The cultured human amniotic fluid-derived mesenchymal stem cells proliferated rapidly after passage. Karyotype mapping showed abnormal female chromosome type with 45, X/46, XX observed. The amniotic fluid-derived mesenchymal stem cells had an immunophenotype similar to that of common mesenchymal stem cells and were positive for CD29, CD44, CD90 and CD105, but negative for CD34 and CD45. The cell cycle measurement showed that amniotic fluid-derived mesenchymal stem cells cultured in vitro could maintain strong proliferation ability. Alkaline phosphatase staining and alizarin red staining results confirmed that amniotic fluid-derived mesenchymal stem cells could be successfully induced to differentiate into osteocytes under specific culture media. These results demonstrated that amniotic fluid-derived mesenchymal stem cells in 45, X/46, XX (Turner’s syndrome) were successfully isolated and the cells had a great potential of proliferation and showed the characteristics of mesenchymal stem cells. Figures and Tables | References | Related Articles | Metrics

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Growth and differentiation of neural stem cells from neurospheres after culture in serum-free medium Zhao Ji-tong, Shen Qiang, Huang Fei 2013, 17 (14):  2592-2596.  doi: 10.3969/j.issn.2095-4344.2013.14.018 Abstract ( 511 )   PDF (1158KB) ( 571 )   Save BACKGROUND: In vitro isolation and culture of neural stem cells with high purity, strong viability and homogeneous biological characteristics are the basis of neural stem cell transplantation. OBJECTIVE: To establish a method of isolation, cultivation and purification of rat neural stem cells in vitro, to observe cell morphology, and to assess surface markers and multi-directional differentiation capacity. METHODS: Neural stem cells were isolated from rat embryonic cerebral cortex and cultured in serum-free medium. Immunocytochemical staining was used to examine the markers of neural stem cells and their differentiated cells. Cell surface markers were assessed by flow cytometry. RESULTS AND CONCLUSION: Neural stem cells grew as free-floating neurospheres in the serum-free conditions.This method is simple and can isolate, purify and amplify neural stem cells in vitro. The obtained cells have the general biological characteristics of neural stem cells, accounting for 91.5% by flow cytometry at passage 3, and also have multi-directional differentiation potential. Immunocytochemical staining and flow cytometry demonstrated that neural stem cells can differentiate into neurons, astrocytes and oligodendrocytes. Figures and Tables | References | Related Articles | Metrics

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Eupolyphaga sinensis-containing serum influences the adipogenic differentiation of bone marrow mesenchymal stem cells Qi Zhen-xi, Li Shu-qiang, Yu Tao 2013, 17 (14):  2597-2602.  doi: 10.3969/j.issn.2095-4344.2013.14.019 Abstract ( 348 )   PDF (1616KB) ( 551 )   Save BACKGROUND: The experiments try to explain the pathogenesis of Eupolyphaga sinensis for the treatment of avascular necrosis of femoral head through investigating the effect of Eupolyphaga sinensis on the adipogenic differentiation of hormone-induced bone marrow mesenchymal stem cells. OBJECTIVE: To investigate the intervention effect of Eupolyphaga sinensis on the adipogenic differentiation of dexamethasone-induced bone marrow mesenchymal stem cells. METHODS: High-dose dexamethasone was used to induce the adipogenic differentiation of in vitro cultured bone marrow mesenchymal stem cells, and during the adipogenic differentiation, Eupolyphaga sinensis-containing serum was used for intervention. After 6 days intervention, expressions of adipogenic markers, such as triglyceride, peroxisome proliferator-activated receptor mRNA and adipocyte protein 2 mRNA, were detected. RESULTS AND CONCLUSION: Eupolyphaga sinensis could inhibit the expression of peroxisome proliferator-activated receptor mRNA and adipocyte protein 2 mRNA in the differentiation of bone marrow mesenchymal stem cells, and could reduce the level of triglyceride, which suggested that Eupolyphaga sinensis drug can reverse the increased adipogenic differentiation of bone marrow mesenchymal stem cells caused by high-dose hormones.  Figures and Tables | References | Related Articles | Metrics

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Osteogenic characteristics of Naringin-induced rabbit bone marrow mesenchymal stem cells Yang Yuan, Li Xiao-feng, Luo Dao-ming, Wen Chao-hai 2013, 17 (14):  2603-2608.  doi: 10.3969/j.issn.2095-4344.2013.14.020 Abstract ( 278 )   PDF (825KB) ( 435 )   Save BACKGROUND: Drynaria can promote bone growth and has made good clinical curative effect. Can the main ingredients of Naringin induce bone marrow measenchymal stem cells to differentiation into osteoblasts? OBJECTIVE: Through the use of Naringin to induce osteogenic differentiation of rabbit bone marrow mesenchymal stem cells, to observe the osteogenic ability of Naringin-induced rabbit bone marrow mesenchymal stem cells. METHODS: Adherence screening method was used to separate and culture the rabbit bone marrow mesenchymal stem cells. The well-grown third generation of bone marrow measenchymal stem cells were induced with 50 ug/L Naringin and classic osteogenic inducer respectively to differentiate into the osteoblasts, and the osteogenesis identification was performed. RESULTS AND CONCLUTION: Both the classic osteogenic inducer and Naringin induced liquid could induce the bone marrow mesenchymal stem cells to differentiation into osteoblasts with increasing matrix secretion and calcium nodules formation. The enzyme-linked immunosorbent assay showed that there was no significant difference in absorbance value of the cells between the Naringin induction group and the classic osteogenic inducer group. There was no significant difference in absorbance value of the cells induced with Naringin induced liquid and L-DMEN culture medium (P > 0.05). The results indicate that Naringin can induce the bone marrow mesenchymal stem cells to differentiate into osteoblasts without obvious toxic effect. Figures and Tables | References | Related Articles | Metrics

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Revealing nourishing kidney drugs in the culture and differentiation of stem cells  Gao Yue-cai, Li Meng, Li Rui-yu, Sun Yan-fu 2013, 17 (14):  2609-2616.  doi: 10.3969/j.issn.2095-4344.2013.14.021 Abstract ( 484 )   PDF (805KB) ( 597 )   Save BACKGROUND: The continuous improvement of stem cell transplantation and understanding of the traditional Chinese medicine theory can strengthen the research regarding the effect of nourishing kidney drugs on the differentiation of stem cells. OBJECTIVE: To investigate the interventional effect of nourishing kidney drugs on stem cell culture and transplantation. METHODS: The relative studies on different categories of single herb or compound nourishing kidney drugs for the proliferation, differentiation and induction of stem cells during stem cell transplantation for the treatment of diseases were collected to analyze the experimental data, and to evaluate the interventional effect of nourishing kidney drugs on the stem cell culture and transplantation based on serum pharmacology. RESULTS AND CONCLUSION: According to the basic research on nourishing kidney drugs and stem cells, and based on the existing new theory, the basic theoretical research was deepened continuously, in order to provide scientific theoretical guidance for the research on nourishing kidney drugs and stem cells. Based on the researches on the proliferation, differentiation and transdifferentiation mechanism of stem cells, the nourishing kidney drugs that could regulate stem cell growth were screened out. Then, combined with the achievement obtained from the clinical application, the single herb or compound nourishing kidney drugs were selected targetedly, and the interventional effect of the drugs on stem cell transplantation in the treatment of major diseases was analyzed. Figures and Tables | References | Related Articles | Metrics

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Research progress in osteogenic differentiation of stem cells induced by estrogen Lü Wei-liang, Liao Yi, Tong Ting-hui 2013, 17 (14):  2617-2624.  doi: 10.3969/j.issn.2095-4344.2013.14.022 Abstract ( 361 )   PDF (693KB) ( 557 )   Save BACKGROUND: Estrogen is closely related to normal bone metabolism in human body and provides favorable conditions for osteogenic differentiation of stem cells. OBJECTIVE: To summarize the application, mechanism of action, safety, efficacy, advantages/disadvantages, and characteristics of estrogen in inducing osteogenic differentiation of various stem cells. METHODS: A computer-based online retrieval of PubMed, CNKI and Wanfang databases was performed for searching papers using the key words “estrogen, bone marrow mesenchymal stem cells, adipose-derived stem cells, embryonic stem cells, osteogenesis” in English and Chinese. RESULTS AND CONCLUSION: Fifty papers were suitable for final analysis. Estrogen is the most important endocrine hormone. It is a safe and effective inducing agent used in the osteogenic differentiation of stem cells, promotes the proliferation and differentiation of stem cells and exerts different actions on stem cells. Estrogen directly influences on the estrogen receptor of the stem cells and also directly influences cell proliferation and differentiation by inducing the production of various cytokines. Nevertheless, there are some uncertainties, including estrogen level, estrogen-target gene expression and transcription on stem cells, and cell-scaffold reconstruction, which should be further studied. Figures and Tables | References | Related Articles | Metrics

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Glucocorticoids promote osteogenic differentiation of mesenchymal stem cells Dong Ping, Xiao Ran 2013, 17 (14):  2625-2632.  doi: 10.3969/j.issn.2095-4344.2013.14.023 Abstract ( 387 )   PDF (814KB) ( 476 )   Save BACKGROUND: Glucocorticoids are immunosuppressive and anti-inflammatory drugs which are widely used in the clinic. However, long-term use of these drugs may cause several side effects, such as osteoporosis. It has been demonstrated that glucocorticoids promote the apoptosis of osteoblasts and osteocytes, and thus inhibit the differentiation of mesenchymal stem cells into osteoblasts, resulting in a decrease number of osteoblasts. However, glucocorticoids can stimulate osteogenic differentiation of mesenchymal stem cells by enhancing the expression of osteoblastic markers under appropriate induction conditions. OBJECTIVE: To review the effect of dexamethasone on the osteogenic differentiation of mesenchymal stem cells and the possible molecular mechanism. METHODS: The PubMed database was retrieved for articles regarding dexamethasone and osteogenic differentiation of mesenchymal stem cells published from 1978 to 2012. The English key words were “mesenchymal stem cell, dexamethasone, osteogenesis, osteoporosis, Runx2, BMP, Noggin, GILZ, glucocorticoid receptor”. A total of 55 articles were suitable for final analysis after repetitive articles were excluded. RESULTS AND CONCLUSION: Dexamethasone controls the osteogenic differentiation of mesenchymal stem cells by regulating the expression of Runx2, Noggin and glucocorticoid-induced leucine zipper. Glucocorticoid receptor may mediate the effect of glucocorticoids, or 11β-hydroxysteroid dehydrogenase interconverts the activity of glucocorticoids before it binds to glucocorticoid receptor. In addition, glucocorticoids can promote the osteogenic differentiation of mesenchymal stem cells at a physiological dose of 10-8 mol/L while it inhibits the osteogenic differentiation at a pharmacological dose of ≥10-7 mol/L. Figures and Tables | References | Related Articles | Metrics

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Autologous endothelial progenitor cells for treatment of ischemic/hypoxic brain injury  Cui Li-ling, Huang Guo-zhi, Chen Zhen-zhou, Guo Yang 2013, 17 (14):  2633-2640.  doi: 10.3969/j.issn.2095-4344.2013.14.024 Abstract ( 331 )   PDF (673KB) ( 468 )   Save BACKGROUND: Following ischemic/hypoxic brain injury, neurogenesis and neurofunctional recovery are closely related to vascular formation and plasticity in ischemic region. Vascular endothelial progenitor cells participate in vascular formation and repair in postnatal ischemic tissue, promote the recanalization of blood flow and the supply of nutritive substances such as oxygen, providing microenvironment for neurofunctional recovery. OBJECTIVE: To evaluate the feasibility, efficacy and safety of use of autologous vascular endothelial progenitor cells in the treatment of ischemic/hypoxic brain injury and investigate a new method for improving the neurological function of patients with ischemic/hypoxic brain injury. METHODS: A computer-based online retrieval of PubMed, ScienceDirect, Springerlink and CNKI databases was performed for papers describing use of vascular endothelial progenitor cells in the treatment of ischemic/hypoxic brain injury using the key words “EPCs, endothelial progenitor cell, stroke” in English and Chinese. In the same research filed, papers that published recently or in high impact factor journals were selected. A total of 43 papers were suitable for final analysis. RESULTS AND CONCLUSION: Following ischemic/hypoxic brain injury, neurogenesis and neurofunctional recovery are closely related to vascular formation and plasticity in ischemic region. Vascular endothelial progenitor cells participate in vascular formation and repair in postnatal ischemic tissue, promote the recanalization of blood flow and the supply of nutritive substances such as oxygen, providing microenvironment for neurofunctional recovery. The use of autologous vascular endothelial progenitor cells in the treatment of ischemic/hypoxic brain injury is feasible, safe and effective. Nevertheless, a larger number of biological and animal experiments are needed for providing theoretical evidence for clinical application of autologous vascular endothelial progenitor cells. Figures and Tables | References | Related Articles | Metrics

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Isolation and identification of osteosarcoma stem cells  Qu Shao-zheng, Li Shu-zhong, Gao Jia-ke, Zhang Jin-feng 2013, 17 (14):  2641-2648.  doi: 10.3969/j.issn.2095-4344.2013.14.025 Abstract ( 392 )   PDF (686KB) ( 534 )   Save BACKGROUND: Osteosarcoma is the most common bone primary malignant tumors, and the occurrence of osteosarcoma may relate with osteosarcoma stem cells. OBJECTIVE: To evaluate the isolation, culture and identification method of osteosarcoma stem cells, to investigate the expression of relative tumor markers. METHODS: The osteosarcoma stem cells were isolated and cultured with magnetic activated cell sorting method under serum-free condition and serum-free joint antineoplastic condition, in order to sort Stro-1 positive and CD133 positive osteosarcoma stem cells. The expression level and tumorigenic properties of CD133, Oct3 / 4 and Nanog markers of osteosarcoma cancer stem cells were tested with immunofluorescence staining and Western blotting methods. RESULTS AND CONCLUSION: Suspended cell condensation could be seen in the osteosarcoma stem cells after culture for 2-10 days, the proliferation incubation period was about 24 hours, and the Stro-1 positive stem cells could form the suspended cell condensation, while the Stro-1 negative stem cells could not form the suspended cell condensation. In addition, osteosarcoma stem cells could highly express Oct3/4, Nanog and CD133, the CD133-positive osteosarcoma stem cells could highly express CD133 molecule with strong invasiveness, while the CD133 negative cells could not express CD133 molecule and the invasiveness was weak. Celecoxib could inhibit the formation of osteosarcoma stem cells to some extent, and could reduce the expression of tumor angiogenic blood vessels and vascular endothelial growth factor. Figures and Tables | References | Related Articles | Metrics

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Isolation and culture of mesenchymal stem cells: From laboratory research to clinical application   Li Bing-yao, Wu Xiao-yun, Wu Yan1 2013, 17 (14):  2649-2655.  doi: 10.3969/j.issn.2095-4344.2013.14.026 Abstract ( 1080 )   PDF (636KB) ( 1130 )   Save BACKGROUND: Mesenchymal stem cells are multipotent adult stem cells. The source, isolation and culture of mesenchymal stem cells differ greatly in different studies in terms of concepts, research methods and outcomes. OBJECTIVE: To investigate the progress in isolation and culture of mesenchymal stem cells. METHODS: The PubMed database (http://www.ncbi.nim.nih.gov/PubMed) was retrieved for articles regarding application of stem cells from 2007 to 2011 by computer. The English key words are “mesenchymal stem cells” and “source isolation or culture”. The papers published recently or in authoritative journals were selected. Forty-three papers were suitable for final analysis.   RESULTS AND CONCLUSION: A standardized culture system should be established to achieve isolation and expansion in vitro of mesenchymal stem cells. Therefore, basic research of mesenchymal stem cells should be further developed, and the system of in vitro expansion and oriented differentiation of mesenchymal stem cells should be established, and study on the effects of transplantation of gene-transfected mesenchymal stem cells is of great significance. Figures and Tables | References | Related Articles | Metrics

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Mesenchymal stem cell transplantation for treatment of myocardial infarction:Transplantation strategy and efficacy evaluation Chen Ling-ling, Yin Li-xue 2013, 17 (14):  2656-2660.  doi: 10.3969/j.issn.2095-4344.2013.14.027 Abstract ( 285 )   PDF (540KB) ( 513 )   Save BACKGROUND: Numerous studies have demonstrated that mesenchymal stem cells exhibit great potential in treatment of myocardial infarction. OBJECTIVE: To review the strategies of mesenchymal stem cell transplantation for treatment of myocardial infarction and evaluate the therapeutic effects after cell transplantation. METHODS: A computer-based online retrieval of PubMed, CNKI, CQVIP and FMJS databases was performed to search papers regarding therapeutic effects of mesenchymal stem cell transplantation with key words “mesenchymal stem cell; function, angiogenesis or survival” in Chinese and English. Papers describing different transplantation methods to enhance heart function recovery, angiogenesis and survivalrate after mesenchymal stem cell transplantation were searched. In the same research field, papers published recently or in high-impact journals were selected. A total of 220 papers were acquired in the initial retrieval. According to inclusion criteria, 20 papers were suitable for final analysis. RESULTS AND CONCLUSION: Stem cell in vitro expansion and differentiation followed by transplantation have emerged as a promising alternative therapeutic approach for cardiovascular disease, but existing strategies are restricted by low cell engraftment, differentiation and survival. In addition, it is also important that reasonable methods should be adopted to accurately assess transplantation efficacy for determining later transplantation strategies. Heart function reconstruction, myocardial fibrosis inhibition and angiogenesis are mainly used as evaluation criteria. From the global to the local function and from the right single imaging to “multimodality noninvasive imaging” are two new developments. How to choose appropriate transplantation strategies and efficacy evaluation method to enhance the efficacy of mesenchymal stem cell therapy needs further studies. Figures and Tables | References | Related Articles | Metrics