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05 March 2014, Volume 18 Issue 10 Previous Issue    Next Issue

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Paracrine effects of bone marrow mesenchymal stem cells on biological function of osteoblasts  Li Cheng, Zhou Hai-bin 2014, 18 (10):  1477-1483.  doi: 10.3969/j.issn.2095-4344.2014.10.001 Abstract ( 368 )   PDF (1258KB) ( 595 )   Save BACKGROUND: Some studies have shown that bone marrow mesenchymal stem cells have a certain role in the repair of bone defects, but the clinical application is limited because of ischemia, hypoxia and inflammation of injured tissues. OBJECTIVE: To investigate the paracrine effects of bone marrow mesenchymal stem cells on osteoblast biological function. METHODS: Bone marrow mesenchymal stem cells were isolated using standard Ficoll-Paquedensity gradient centrifugation. Mesenchymal stem cell conditioned medium was prepared to cultivate osteoblasts, MG63. Proliferation of MG63 cells was analyzed by cell counting kit-8. Migration of MG63 cells was analyzed by cell scratch method. Alkaline phosphatase activity of MG63 cells was analyzed by microplate test kit. Real-time PCR was performed to evaluate osteoblast differentiation markers, alkaline phosphatase, collagen type I and osteocalcin. Alizarin red staining was performed to evaluate osteoblast mineralization. RESULTS AND CONCLUSION: The cells were strongly positive for CD44, CD73 and CD90, but negative for CD34. MG63 cells cultured in the conditioned medium showed better proliferation and migration than those cultured in the Dulbecco’s modified Eagle’s medium. The activity and mRNA expression of alkaline phosphatase  were much higher after induction of 4, 7 days (P < 0.01). There was no significant difference in expression of collagen type I and osteocalcin after induction of 4 days, but they were significantly higher than those in the control group after induction of 7 days (P < 0.05). Alizarin red staining showed that the number of calcium nodules was increased and the mineral apposition was enhanced after induction of 21 days with the conditioned medium. These findings suggest that the paracrine substance of bone marrow mesenchymal stem cells can significantly promote osteoblast proliferation, migration, differentiation and mineralization. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Biological characteristics and superiority of rat bone marrow mesenchymal stem cells isolated and cultured using whole bone marrow adherence method Li Shuang-yue, Qi Yuan, Chen Ruo-lin, Wang Zhe-min, Liu Shuang, Piao Feng-yuan 2014, 18 (10):  1484-1489.  doi: 10.3969/j.issn.2095-4344.2014.10.002 Abstract ( 326 )   PDF (563KB) ( 456 )   Save BACKGROUND: Bone marrow mesenchymal stem cells are rare in vivo. It is important to purify, proliferate and differentiate bone marrow mesenchymal stem cells in vitro for further research. OBJECTIVE: To evaluate the biological characteristics, phenotype and multiple differentiation potential cultivation of bone marrow mesenchymal stem cells that are isolated, cultured and purified using the whole bone marrow adherence method. METHODS: Bone marrow mesenchymal stem cells were isolated, purified and cultured by the whole bone marrow adherence method. Morphological observation and flow cytometry determination of cell surface markers were performed. Osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was induced. RESULTS AND CONCLUSION: We successfully purified and proliferated bone marrow mesenchymal stem cells with high cell viability and differentiation ability. Fibroblast-like cells were harvested, expressing CD29 and CD90, but not CD45. Following osteogenic and adipogenic induction, cells were positive for oil red O staining and alizarin red staining. The whole bone marrow adherence method is easy to operate, has little impact on cell viability, and can be used to harvest high-purification bone marrow mesenchymal stem cells with high cell viability and differentiation ability. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human synovial fluid promotes directed differentiation of bone marrow mesenchymal stem cells Hua Qiang, Wu Jia-qi, Zhong Chuan-shan, Liu Zong-chao, Yan Guang-jian, Xiong Xiao-tian, Cui Xiao-ming 2014, 18 (10):  1490-1495.  doi: 10.3969/j.issn.2095-4344.2014.10.003 Abstract ( 355 )   PDF (601KB) ( 436 )   Save BACKGROUND: Nowadays, growth factors are commonly used to induce bone marrow mesenchymal stem cells. However, this is a high-cost method with a great amount of growth factors. In addition, the chondrogenic potential of bone marrow mesenchymal stem cells will decrease significantly with increasing times of culture. OBJECTIVE: To observe the directed differentiation of bone marrow mesenchymal stem cells co-cultured with human synovial fluid. METHODS: Human bone marrow mesenchymal stem cells were isolated and cultured by adherence screening method. The synovial fluid of the knee was aspirated from healthy volunteers by aseptic operation. Passage 3 human bone marrow mesenchymal stem cells were co-cultured with the following media: synovial fluid+complete medium; synovial fluid+bone marrow mesenchymal stem cells+complete medium; bone marrow mesenchymal stem cells+complete medium. The morphology and growth of the cells were observed under an inverted microscope every day. At days 7, 14 and 21 of induction, toluidine blue staining and immunocytochemical staining were performed. RESULTS AND CONCLUSION: After co-culture with human synovial fluid, human bone marrow mesenchymal stem cells proliferated slowly, and varied from fusiform to oval or polygonal; toluidine blue and collagen II staining were positive. These findings indicate that the synovial fluid has a positive role in the chondrogenic differentiation of bone marrow mesenchymal stem cells. The synovial fluid may contain substances that promote the chondrogenic differentiation of bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of Zuoguiwan and Youguiwan on transforming growth factor beta1 and its signal transduction protein Smad2/3 in osteogenic induction of rat bone marrow mesenchymal stem cells  Sun Yue-jiao, Song Nan, He Wen-zhi, He Li-juan, Ren Yan-ling 2014, 18 (10):  1496-1501.  doi: 10.3969/j.issn.2095-4344.2014.10.004 Abstract ( 335 )   PDF (501KB) ( 552 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE: To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factor β1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cell cultivation was processed in five groups: bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, and β-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I collagen. The immunohistochemical assay was utilized to test transforming growth factor β1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION: It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I collagen, transforming growth factor β1 and Smad2/3 expression, compared with blank serum group and inducer group (P < 0.05); moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P < 0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Wnt signaling pathway by which puerarin suppresses adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells Qi Zhen-xi, Zhang Zhan-yong, Wan Tian, Wu Min-rui, Chen Han-yao 2014, 18 (10):  1502-1507.  doi: 10.3969/j.issn.2095-4344.2014.10.005 Abstract ( 365 )   PDF (475KB) ( 509 )   Save BACKGROUND: Recently, glucocorticoid-induced necrosis of femoral head has been much studied. However, the precise Wnt signaling pathway by which puerarin suppresses adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells remains unconfirmed. OBJECTIVE: To investigate the expression of Wnt signaling pathway related genes and the key factor protein, β-catenin, during adipogenic differentiation of glucocorticoid-induced rat bone marrow mesenchymal stem cells treated by puerarin. METHODS: The third generation of bone marrow mesenchymal stem cells were cultured with Dulbecco’s modified Eagle’s medium containing blank serum (blank control group), dexamethasone (hormone group), dexamethasone with puerarin low dose group, the middle dose group and high dose group. After 6 days of culture, in the above five groups, the expressions of Wnt/β-catenin signaling pathway members, Wnt10b mRNA, GSK3β mRNA, β-catenin mRNA, were detected using RT-PCR assay, and the expression of β-catenin protein was detected using western blot assay.  RESULTS AND CONCLUSION: Compared with the control group, the Wnt10b mRNA, β-catenin mRNA and β-catenin protein expressions were significantly higher in puerarin groups, but GSK3β mRNA expression was significantly lower in the puerarin groups. These findings suggest that puerarin effects on inhibition of adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells probably are realized through the activation of Wnt/β-catenin signal pathway and regulation of the key factor Wnt10b mRNA, GSK3β mRNA, β-catenin mRNA, and β-catenin protein expressions. The mechanism by which puerarin prevents glucocorticoid-induced necrosis of femoral head not only improves local microcirculation of the femoral head, but also relates to its inhibitory effects on adipogenic differentiation of glucocorticoid-induced bone marrow mesenchymal stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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26RFa effects on osteogenic differentiation of human bone marrow mesenchymal stem cells  Du Bin, Lin Qing, Liu Meng-jun, Chen Zhi-xin 2014, 18 (10):  1508-1513.  doi: 10.3969/j.issn.2095-4344.2014.10.006 Abstract ( 301 )   PDF (474KB) ( 372 )   Save BACKGROUND: Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism. OBJECTIVE: To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells.                    METHODS: In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. Cells were inoculated into 6-well plates and then divided into two groups: experimental group treated with 10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay. RESULTS AND CONCLUSION: After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group (P < 0.05, P < 0.01, P < 0.05). After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group (P < 0.05). These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Histone deacetylases mRNA profile in mesenchymal stem cells derived from ovariectomized mice Liu Da-yong, Tai Yong, Wang Mei-rui, Cui Ting, Liu Ping, Zhao Meng-ming, Jia Zhi 2014, 18 (10):  1514-1520.  doi: 10.3969/j.issn.2095-4344.2014.10.007 Abstract ( 308 )   PDF (496KB) ( 508 )   Save BACKGROUND: As the multipotent differentiation potential, bone marrow mesenchymal stem cells exert an important role in bone metabolism disorders, which is regulated by a variety of hormones and cytokines. Currently, the epigenetic regulatory mechanisms underlying osteogenic differentiation of bone marrow mesenchymal stem cells are unclear, and association between histone deacetylase (Hdac) and osteoporosis needs to be further explored. OBJECTIVE: To investigate the epigenetic mechanisms of bone formation by analyzing the expression of Hdac1, 3, 4 mRNA profile in bone marrow mesenchymal stem cells isolated from ovariectomized mice. METHODS: A total 30 female healthy Kunming mice were randomly divided into sham group and ovariectomy group. After 7 days of adaptive feeding, mice in the ovariectomized group (n=15) were subject to bilateral ovariectomy; mice in sham group (n=15) were sham-operated. RESULTS AND CONCLUSION: In the ovariectomy group, the trabecular bone of the femur was sparse or broken, the width of trabecular bone was narrowed, the trabecular separation was widened, and the trabecular bone volume was reduced. The level of Hdac3 mRNA was lower in bone marrow mesenchymal stem cells from ovariectomized mice compared with controls, but there was no significance between Hdac1, Hdac4 mRNA expression of the two groups. These findings demonstrate that Hdac might play an important role in bone remodeling in the model of estrogen deficiency-induced osteoporosis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Large-scale expansion of clinical-grade human adipose-derived stem cells using the extracellular matrix Su Yue-han, Wei Chao, Lv Pin-lei, Cao Yun, Qiu Yun, Zheng Qing, Xiao Shu-dong, Wang Zheng 2014, 18 (10):  1521-1531.  doi: 10.3969/j.issn.2095-4344.2014.10.008 Abstract ( 395 )   PDF (1047KB) ( 915 )   Save BACKGROUND: Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE: To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS: In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro  expansion, total cell number, expression of cell surface markers, cell senescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION: Total cell number of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cell surface markers. Cellular senescence examination showed that almost all of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could still differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. All above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Immunoregulation role of adipose mesenchymal stem cells in J774.1 cells Bai Li, Zhang Wei, Pang Chun-yan, Wei Fei-fei, Wang Yong-fu 2014, 18 (10):  1532-1538.  doi: 10.3969/j.issn.2095-4344.2014.10.009 Abstract ( 397 )   PDF (630KB) ( 599 )   Save BACKGROUND: Recent studies have found that adipose mesenchymal stem cells can inhibit immune responses of T cells, B cells and dendritic cells. But it is unclear whether adipose mesenchymal stem cells are able to regulate the immune responses of macrophages. OBJECTIVE: To observe the expression of interleukin-6 and tumor necrosis factor α in J774.1 cells co-cultured with adipose mesenchymal stem cells co-cultured. METHODS: Adipose mesenchymal stem cells at a density of 2×107 per well were inoculated into the upper chamber of the transwell and 24-well plates. Then, J774.1 cells were suspended with cell culture medium containing 1 mg/L lipopolysaccharide and planted into the lower chamber of the transwell and 24-well plates containing adipose mesenchymal stem cells as co-culture group. Meanwhile, inactive J774.1 cells to lipopolysaccharide-activat J774.1 cells served as negative and positive control groups, respectively. After 48 hours, the J774.1 cells were collected to extract the RNA samples. Then, mRNA expressions of interleukin-6 and tumor necrosis factor α were measured by real-time quantitative PCR. RESULTS AND CONCLUSION: Compared with the positive control group, direct-contact co-culture of  adipose mesenchymal stem cells and J774.1 cells significantly reduced expressions of interleukin-6 and tumor necrosis factor α in J774.1 cells, but non-contact co-culture only reduced interleukin-6 expression. These findings indicate that adipose mesenchymal stem cells can inhibit the immune response of J774.1 cells activated by lipopolysaccharide. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Large-scale expansion of human umbilical cord mesenchymal stem cells in human platelet lysate as a substitute of fetal bovine serum Li Bing-yao, Wu Xiao-yun, Wu Yan 2014, 18 (10):  1539-1546.  doi: 10.3969/j.issn.2095-4344.2014.10.010 Abstract ( 435 )   PDF (615KB) ( 1222 )   Save BACKGROUND: Classic media and fetal bovine serum are commonly used in the culture of umbilical cord mesenchymal stem cells, but the potential risk of serum culture limits its clinical application. OBJECTIVE: To use human platelet lysate alternative to fetal bovine serum for large-scale production of mesenchymal stem cells for therapeutic applications. METHODS: Human platelet lysate was prepared by repeated freezing and thawing, centrifugation, filtration, and concentration. Human umbilical cord mesenchymal stem cells were cultured in Iscove’s Modified Dulbecco’s Medium containing 5% concentrated platelet lysate (experimental group) or 10% fetal bovine serum (control group). After separation and enzymatic digestion, human umbilical cord mesenchymal stem cells were subcultured at a concentration of 3 000/cm2 up to the fifth generation. Then, cell morphology and diameter, immune phenotype, osteogenic and adipogenic differentiation, and cloning efficiency were detected and compared between two groups. RESULTS AND CONCLUSION: Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a smaller size and more elongated morphology than those in fetal bovine serum-supplemented media. Colony forming unit-fibroblast analyses further showed no significant differences in colony efficiency. Human umbilical cord mesenchymal stem cells cultivated in human platelet  lysate-supplemented media showed an increase of proliferation capacity; whereas, similar immunophenotypes remained in the two groups. In vitro assays revealed intact differentiation potential. Moreover, human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a significantly higher capacity to differentiate towards osteocytes, indicating human platelet lysate is an alternative to fetal bovine serum for low-density production of mesenchymal stem cells for therapeutic applications. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of Corning® CellBIND® Surface medium on growth of human umbilical cord mesenchymal stem cells Wan Ying, Ben Liang, Guan Zi-qiu, Li Chao, Zhang Shi-dong, Nie De-zhi 2014, 18 (10):  1547-1553.  doi: 10.3969/j.issn.2095-4344.2014.10.011 Abstract ( 263 )   PDF (766KB) ( 646 )   Save BACKGROUND: Stem cells expansion technology in vitro is affected by the microenvironment of the culture system. So, to find an effective method is particularly important to promote cell adherent and growth. OBJECTIVE: To compare the effects of different culture media on cell expansion. METHODS: Human umbilical cord mesenchymal stem cells at a density of 5.0×104 cells/cm2 were inoculated into Corning® polystyrene culture dishes coated with or without poly-L-ornithine and Corning® CellBIND culture dishes. Cell adhesion and proliferation were observed, and expressions of cell adhesion proteins and cell markers were detected. RESULTS AND CONCLUSION: Cell adhesion was promoted when cells were cultured in Corning® CellBIND® Surface medium coated with poly-L-ornithine for 24 hours, and the cultured cells grew at the logarithmic phase. The cell proliferation was also enhanced, and the cells expressed cell adhesion protein but not the cell markers of CD73, CD90, CD105. Corning® CellBIND® Surface medium was superior to Corning® polystyrene culture medium in the improvement of cell adhesion and proliferation. Additionally, both of these two media showed no influence on cell phenotype. These findings indicate that Corning® CellBIND® Surface medium can promote cell adhesion and proliferation, but shows no effects on cyclin and cell phenotype. This medium coated with poly-L-ornithine can further accelerate cell adhesion and proliferation, and stably express cell phenotype of human umbilical cord mesenchymal stem cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Human umbilical cord mesenchymal stem cells are induced in vitro to differentiate into fibroblasts Yang Yi, Luo Xin, Jiang Xue-feng, Shuai Han-lin, Song Hong, Zhang Jing-li, Lan Jian-fa 2014, 18 (10):  1554-1559.  doi: 10.3969/j.issn.2095-4344.2014.10.012 Abstract ( 646 )   PDF (626KB) ( 768 )   Save BACKGROUND: The umbilical cord mesenchymal stem cells possess multipotent differentiation capacity, but less research focus on its differentiation into fibroblasts. OBJECTIVE: To investigate the capacity of human umbilical cord mesenchymal stem cells differentiating into fibroblasts. METHODS: Using adherent method, human umbilical cord mesenchymal stem cells were isolated, and flow cytometric analysis of the surface antigen was performed. Passage 3 cells were selected for osteogenic and adipogenic differentiation, and cells differentiated into fibroblasts under the induction of basic fibroblast growth factor. RESULTS AND CONCLUSION: Adherent stem cells were stably isolated from the umbilical cord. Human umbilical cord mesenchymal stem cells lowly expressed CD31, CD45, CD40, HLA-DR, but strongly expressed CD29, CD90, CD44, CD105. Oil red O staining showed red droplets were full of the cytoplasm after adipogenic induction; alizarin red staining showed red calcium nodules after osteogenic induction. After induced by basic fibroblast growth factor, the type I collagen expression was significantly higher than that in the control group. These findings indicate that the adherent human umbilical cord mesenchymal stem cells are reliably isolated with high purity; basic fibroblast growth factor can induce differentiation of umbilical cord mesenchymal stem cells into fibroblasts. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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MRI tracking of superparamagnetic iron oxide-labeled rabbit bone marrow mesenchymal stem cells for treatment of femoral head avascular necrosis Meng Zeng-dong, Li Lei, Hu Biao, Lei Yun-kun, Liu Wei, Tang Xu 2014, 18 (10):  1560-1565.  doi: 10.3969/j.issn.2095-4344.2014.10.013 Abstract ( 379 )   PDF (807KB) ( 495 )   Save BACKGROUND: Bone marrow mesenchymal stem cell (BMSC) transplantation is one of the developmental directions in the treatment of femoral head necrosis. In recent years, the use of superparamagnetic iron oxide (SPIO) nanoparticles labeled target cells traced by MRI imaging method has become the focus of the study. OBJECTIVE: To observe the in vivo MRI tracking and the curative effects of SPIO-labeled BMSC transplantation on rabbit femoral head necrosis. METHODS: SPIO-labeled BMSCs, unlabeled BMSCs, and normal saline were injected in situ into the necrotic femoral head of rabbits. Following MRI dectection, the image changes of transplanted BMSCs marked by SPIO were observed among the three scanning sequences of SE T2WI, FSE T2WI and GRE T2*WI. Meanwhile, the area percentage of newly formed bone trabecula in the defect samples under high power lens were observed and calculated for statistical analysis. RESULTS AND CONCLUSION: In situ cell transplantation group showed the emerging and extinctive time of the decreased-signal region was different among the three scanning sequences of SE T2WI, FSE T2WI and GRE  T2*WI. It was found that the decreased-signal region of the MRI scanning sequences was the target of the present experiment. No obvious signal change occurred in the control side. After 6 weeks of transplantation, the area percentage of newly formed bone trabecula in the defect samples showed no difference in SPIO-labeled and unlabeled BMSC transplantation groups (P > 0.05), but it was higher than that in the control side (P < 0.01). The SPIO-labeled BMSCs and unlabeled BMSCs are shown to have the same effects in the treatment of femoral head necrosis. The SPIO-labeled BMSCs can be observed obviously by MRI detection in vitro. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effects of intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells on myocardial cardiac troponin and serum-related factors in rats Wang Si-ping, Nie Na-na, Wang Li, Li Zi-pu 2014, 18 (10):  1566-1572.  doi: 10.3969/j.issn.2095-4344.2014.10.014 Abstract ( 270 )   PDF (515KB) ( 460 )   Save BACKGROUND: Current researches on whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells (UC-MSCs) is safe or not lack theoretical basis. OBJECTIVE: To explore the effects of intramuscular injection of heterogeneous UC-MSCs on expression of myocardial vascular endothelial growth factor (VEGF), cardiac troponin I (cTnI) and serum VEGF, hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1) and granulocyte macrophage colony stimulating factor (GM-CSF) in normal Wistar rats. METHODS: A total of 60 Wistar rats were divided into six groups randomly. Rats in the six groups were respectively administrated with intramuscular injection of PBS, Dulbecco’s modified Eagle’s medium, hUC-MSC supernatant, 0.25×105, 1.0×105, 4.0×105 hUC-MSCs. Rats received a second intramuscular injection 4 weeks after first injection. Eight weeks later, the blood sample and myocardial tissue were taken. The serum concentration of VEGF, HGF, IGF-1 and GM-CSF were measured with enzyme-linked immunosorbent assay kit. The myocardial VEGF and cTnI expression were detected by immunohistochemical technique.  RESULTS AND CONCLUSION: There was no significant difference in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF among the groups before and after injection (P > 0.05). In the same group, no statistical significant changes in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF were found among the rats before and after injection (P > 0.05). Eight weeks after injection, weak positive expression of the VEGF in the cytoplasm of cardiocytes in the six groups was observed, and strong positive expression of the cTnI in the cytoplasm of cardiocytes in the six groups was observed. There was no significant difference in the VEGF and cTnI content in the myocardium among the groups (P > 0.05). Intramuscular injection of hUC-MSCs or the supernatant of hUC-MSCs had no effects on the serum concentration of the VEGF, HGF, IGF-1, GM-CSF and the myocardial VEGF and cTnI expression in normal Wistar rats. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Effect of basic fibroblast growth factor on the migration of human adipose-derived stem cells toward vascular endothelium Zhu Meng-lin, Jiang Nan, Xu Yang-yang, Cao Jing, Yang Liu 2014, 18 (10):  1573-1578.  doi: 10.3969/j.issn.2095-4344.2014.10.015 Abstract ( 369 )   PDF (498KB) ( 578 )   Save BACKGROUND: The establishment of a good blood supply is a key mechanism for successful implantation of engineered tissues. OBJECTIVE: To observe the effect of basic fibroblast growth factor on the migration of human adipose-derived stem cells via implanting the human adipose-derived stem cells and sodium hyaluronate composite graft at the subcutaneous site of BALB/C mice, in order to explore an optimal scheme for soft tissue reconstruction. METHODS: Human adipose-derived stem cells were isolated from the adipose tissue of healthy cosmetic patients which received liposuction, and the cells were subcultured. Then 5×109/L passage 3 cell suspension labeled by cm-dil was prepared. The working solution containing 2 mg/L basic fibroblast growth factor was prepared. Composite tissue al-lografts which were the mixtures of 0.25 mL sodium hyaluronate, 0.2 mL cell suspension and 0.05 mL working solution or DMEM were implanted into the subcutaneous site of both sides of the mouse back. Specimens were taken at 6 weeks after operation and were evaluated histologically after hematoxylin-eosin and vascular immunofluorescent staining. RESULTS AND CONCLUSION: No necrosis, liquefaction, nodular tissue or gel remained in operated position. The hematoxylin-eosin staining showed the main components of the specimens were the adipose tissue and the  loose connective tissue. The immunofluorescence staining showed the overlaps between the cm-dil fluorescence from human adipose-derived stem cells and the FITC fluorescence from the vascular endothelium in the experimental group were more than those in the control group (P < 0.05). Basic fibroblast growth factor promotes the migration and the differentiation of human adipose-derived stem cells in the sodium hyaluronate scaffold into vascular endothelium. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Hypoxia-inducible factor-1 alpha effects on bone marrow mesenchymal stem cell mobilization in rats with acute myocardial infarction Qi Jin-wei, Cheng Jing-lin, Zhou Shu, Li Jing-rong, Li Xue-xiang, Yang Qin, Zhang Hao, Wan Jun, Wang Yu-lin, Zhang Li-xin, Chen Yun-yun, Xi Xiu-xia, Ye Li, Tang Qian, Xu Feng, Jang Yang 2014, 18 (10):  1579-1584.  doi: 10.3969/j.issn.2095-4344.2014.10.016 Abstract ( 284 )   PDF (383KB) ( 534 )   Save BACKGROUND: Increasing autologous stem cell mobilization is conceived to achieve effectively repair of cardiac ischemic injury. Therefore, it is important to seek a specific and effective mobilization agent. OBJECTIVE: To observe the effects of hypoxia-inducible factor-1α (HIF-1α) on bone marrow mesenchymal stem cell mobilization in myocardial infarction. METHODS: Left anterior descending artery was ligated to establish a rat model of acute myocardial infarction in 90 outbreeding Sprague-Dawley rats, and then the models were randomly divided into three groups. In HIF-1α-antisense oligonucleotide (ASODN) group, HIF-1α-ASODN was infused into the tail vein to restrain the expression of HIF-1α in infarcted ischemic tissue. In HIF-1α-missense oligonucleotide (MSODN) group or control group, an equal volume of HIF-1α-MSODN or saline was injected. RESULTS AND CONCLUSION: After 30 hours and 7 days of modeling, the number of bone marrow mesenchymal stem cells and expression of vascular endothelial growth factor in the peripheral blood of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group.  After 7 days of modeling, the expressions of HIF-1α protein, vascular endothelial growth factor protein and mRNA in the ischemic myocardial tissues of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. After 7, 14 and 28 days of modeling, the capillary density in the ischemic myocardial tissues of the control group was similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. These findings indicate that after acute myocardial infarction, high expression of HIF-1α exhibits a causal relationship with mobilization of bone marrow mesenchymal stem cells, initiating a series of self-healing process of myocardial tissues. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Creation and effectiveness of mammary stem cell medium Zhang Jun-hong, Yang Jing, Dou Xiao-wei, Wang Chun-hua, Li Qing, Zhao Chun-hua 2014, 18 (10):  1585-1590.  doi: 10.3969/j.issn.2095-4344.2014.10.017 Abstract ( 291 )   PDF (508KB) ( 415 )   Save BACKGROUND: Now in mammary stem cell research, no proper mammary stem cell medium is provided to culture mammary stem cells. OBJECTIVE: To create a mammary stem cell medium and validate its application by isolating Sca-1+ mammary stem cells. METHODS: We first used BM medium to culture mammary organoids, and after 6 days, the expression of Sca-1 and vimentin was detected in fibroblasts by immunofluoresence method. Then, we established MaECM medium which arrested fibroblasts growth. After 6 days culture of mammary organoids by MaECM medium, Sca-1+ and  Sca-1- cell populations were sorted out by magnetic sorting and the purity was analyzed by flow cytometry. Sorted 1×104 Sca-1+ or Sca-1- cells were transplanted into the bilateral mammary fat pads of four mice, and after 6-8 weeks, the fat pads were harvested for whole-mount immunohistochemical analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION: After 6 days culture of mammary organoids under BM medium, small-sized colonies were generated around lots of fibroblasts. Immunofluoresence staining detected strong expression of vimentin and Sca-1 in fibroblasts, indicating that the BM medium is not suitable to isolate Sca-1+ mammary stem cell. The MaECM medium promoted the proliferation of mammary epithelial cells whereas arrested fibroblasts growth. After 6 days culture of mammary organoids under MaECM medium and magnetic sorting, the flow cytometry showed that the purity of Sca-1+ cell reached 92% and 5% in the Sca-1+ and Sca-1- population, respectively. The results from transplantation test showed that six mammary outgrowths were regenerated out of eight injected fat pads in the Sca-1+ cells transplantation, but in the Sca-1- transplantation population, one mouse died and the other transplants failed to produce outgrowths. We developed the MaECM medium which promoted the proliferation of mammary epithelial cells whereas arrested Sca-1+ fibroblasts growth. Using the medium, we confirmed that Sca-1+ mammary cells have capacity of isolating mammary stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Proliferative abilities of placenta-derived mesenchymal stem cells at different passages Bai Jin-ping, Li Xiu-ying, Li Xue, Yang Qi-wei, Liu Ying-nan, Wang Yi-min 2014, 18 (10):  1591-1596.  doi: 10.3969/j.issn.2095-4344.2014.10.018 Abstract ( 332 )   PDF (388KB) ( 617 )   Save BACKGROUND: The mesenchymal stem cells which are from the placenta have better proliferative potential than the bone marrow mesenchymal stem cells. It is inconclusive, however, whether placenta-derived mesenchymal stem cells at different passages have the same proliferative ability. OBJECTIVE: To analyze the proliferative ability of placenta-derived mesenchymal stem cells. METHODS: Placenta-derived human mesenchymal stem cells were isolated with enzyme digestion, and flow cytometry was used to detect the cell surface markers. After osteogenic and adipogenic induction, cell proliferation detection was carried out using cell counting and BrdU analysis. Cell doubling time was calculated based on formula. The maximum cell passage was determined by long term cell culture until the cell died or differentiated. RESULTS AND CONCLUSION: CD44, CD90 and CD105 were highly expressed in placenta-derived mesenchymal stem cells, while CD14, CD34 and CD45 were negatively expressed. After 21 days of adipogenic induction, oil red O staining showed red fat droplets; after 35 days of osteogenic induction, alizarin red staining showed calcium nodules. Cell counting started from passage 1 to 25, and cell number reached to 3.5×1011 at passage 15. Cell proliferation at passage 5 was stronger with shorter doubling time than that at passages 10 and 15. When cells reached to passage 25, three specimens completely altered their cell morphology with cobblestone structure, and another two specimens lost adherent features. These findings indicate that  placenta-derived mesenchymal stem cells can be long-term subcultured upon to passage 25. Different proliferation capacities are found in different passages of placenta-derived mesenchymal stem cells. The proliferation ability of passage 5 placenta-derived mesenchymal stem cells is stronger than that of passage 10 and 15 cells. Placenta-derived mesenchymal stem cells at passages 0-5 are the important source for stem cell therapy and tissue engineering. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Limb ischemic postconditioning promotes the proliferation of endogenous neural stem cells in rats with cerebral ischemia injury Du Guo-jia, Dang Mu-ren, Tan Ze-ming, Su Ri-qing, Shen Qian, Fang Jia-sheng 2014, 18 (10):  1597-1602.  doi: 10.3969/j.issn.2095-4344.2014.10.019 Abstract ( 272 )   PDF (351KB) ( 600 )   Save BACKGROUND: There are a few reports addressing application of limb ischemic preconditioning in cerebral ischemia-reperfusion injury, but few reports have addressed limb ischemic postconditioning applied in this field. OBJECTIVE: To explore the effects of limb ischemic postconditioning on the proliferation of neural stem cells of rats after cerebral ischemia. METHODS: Focal stroke models were established in rats, and then 30 rat models were randomly divided into experimental group (n=15) and control group (n=15). Remote limb ischemic postconditioning was applied in the experimental group, but not in the control group. 5-Bromo-2-deoxyuridine (BrdU; 50 mg/kg) was injected intraperitoneally into rat models three times a day, and then the rat brains were respectively obtained at days 5, 10 and 15 days in two groups. The amount of BrdU-positive cells was measured by immunohistochemistry method in two groups. RESULTS AND CONCLUSION: Compared with the control group, the experimental group was significantly  decreased in the severity of injury and behavior scores (P < 0.05). At various time points, the number of BrdU-positive cells was obviously greater in the experimental group than the control group (P < 0.05). Results have indicated that limb ischemic postconditioning can improve behavior of rats and promote the proliferation of neural stem cells, thereby showing neuroprotective effects against cerebral ischemia injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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In vitro co-culture of human adipose-derived mesenchymal stem cells and T-lymphocytes from patients with aplastic anemia Wang Liang, Xu Min, Zhang Mu-hua, Xing Jian, Zhao Xia, Han Fang, Liu Guo-qiang 2014, 18 (10):  1603-1608.  doi: 10.3969/j.issn.2095-4344.2014.10.020 Abstract ( 297 )   PDF (463KB) ( 427 )   Save BACKGROUND: Clinical infusion of hematopoietic stem cells and mesenchymal stem cells for treatment of aplastic anemia has been reported. OBJECTIVE: To investigate the effect of human adipose-derived mesenchymal stems cells on the secretion function of T lymphocytes of aplastic anemia patients. METHODS: Human adipose-derived mesenchymal stems cells were extracted from healthy human adipose tissues. T-lymphocytes were harvested from peripheral blood of patients with aplastic anemia by density gradient centrifugation. Human adipose-derived mesenchymal stems cells were co-cultured with T-lymphocytes. The levels of interleukin-2, interleukin-4, interleukin-10 and interferon-γ were detected by enzyme linked immunosorbent assay. T-bet and GATA-3 levels were examined by real-time PCR and western blot. RESULTS AND CONCLUSION: The levels of Th1 type cytokines interferon-γ and interleukin-2 in the co-culture group were significantly lower than those in the T-lymphocyte group (P < 0.05). But the levels of Th2 type cytokines interleukin-4 and interleukin-10 in the co-culture group were significantly higher than those in the T-lymphocyte group (P < 0.05). The T-bet mRNA and protein levels in the co-culture group were significantly lower than those in the T-lymphocyte group, while the GATA-3 mRNA and protein levels were significantly higher in the co-culture group. Human adipose-derived mesenchymal stems cells can mediate an immunoregulation effect on T-lymphocytes of aplastic anemia patients in vitro, which is possibly related with the inhibition of Th1-dominant response due to the disorder of T-bet and GATA-3 gene expression. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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An improved method for isolation of human umbilical cord mesenchymal stem cells Li Yan-qi, Wang Hong-yi, Yao Yao, Liu Jing-jing, Xu Xiao, Zhang Yu, Liu Yang, Wu Chu-tse,Jin Ji-de 2014, 18 (10):  1609-1614.  doi: 10.3969/j.issn.2095-4344.2014.10.021 Abstract ( 547 )   PDF (515KB) ( 727 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells with capabilities for self-renewal and multi-differentiation have attracted widespread attention. OBJECTIVE: To develop an efficient method for isolation and culture of human umbilical cord mesenchymal stem cells, and to analyze the cell biological features. METHODS: Mesenchymal stem cells were isolated and cultured from human umbilical cord by improved tissue cultivation. Immunophenotype and cell cycle were analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated by in vitro osteogenic and adipogenic induction as well. RESULTS AND CONCLUSION: Some fusiform cells crawled out from human umbilical cord tissues after cultivation for 5 days and formed colonies about 10 days later. When the removed tissues were further cultured, more cells appeared again within 2 days and formed colonies after 5 days. The isolated cells exhibited similar morphology of fibroblast-like shape after passage. Furthermore, the cells expressed CD90, CD105, but were negative for the markers of CD34, CD45, HLA-DR. Population doubling time of the cells calculated from the result of MTT was about 50 hours and cell cycle analysis showed that 41.24% cells were in the G2/S phrase. Therefore, the isolated cells had a high prolification ability. In addition, the isolated cells could be induced into osteoblasts   and adipocytes in vitro. In a word, the results of this study demonstrated that the cells from the second tissues culture possessed the biological characteristics of mesenchymal stem cells and more primary umbilical cord mesenchymal stem cells were acquired through the improved method. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation combined with edaravone inhibits neuronal apoptosis after cerebral infarction Ma Hai-yan 2014, 18 (10):  1615-1620.  doi: 10.3969/j.issn.2095-4344.2014.10.022 Abstract ( 278 )   PDF (412KB) ( 479 )   Save BACKGROUND: Several studies have confirmed that bone marrow mesenchymal stem cell transplantation exerts a certain neuroprotective role in cerebral infarction. Edaravone is a novel potent small-molecule hydroxyl radical scavenger, which play a protective role in the brain by eliminating free radicals and inhibiting nerve cell damage after cerebral infarction. OBJECTIVE: To explore the influence of bone marrow mesenchymal stem cells combined with edaravone on expressions of aquaporin-4, Bcl-2, and brain-derived neurotrophic factor after cerebral infarction in rats. METHODS: Eighty Wistar rats were selected to establish models of cerebral infarction by right middle cerebral artery occlusion, and then randomly divided into control group, bone marrow mesenchymal stem cells group, edaravone group and edaravone+bone marrow mesenchymal stem cells group (combination group). After 6 hours of modeling, rats from these four groups were respectively injected via the tail vein with PBS, bone marrow mesenchymal stem cells, edaravone and edaravone+bone marrow mesenchymal stem cells. After 72 hours, the  rats were decapitated to take brain tissues. Consequently, expressions of aquaporin-4, Bcl-2, and brain-derived neurotrophic factor mRNA and protein were determined by RT-PCR and western blot methods. After 12, 24 and 36 hours, TUNEL method was used for the determination of cell apoptosis. RESULTS AND CONCLUSION: The expressions of Bcl-2 and brain-derived neurotrophic factor were higher in the combination group than the other three groups (P < 0.05), but the expression of aquaporin-4 was lowest in the combination group (P < 0.05). The number of apoptotic cells in the combination group was significantly lower than that in the other three groups. These findings suggest that the combination of bone marrow mesenchymal stem cells and edaravone can improve expressions of Bcl-2 and brain-derived neurotrophic factor in the injured site, and significantly inhibit neuronal apoptosis. Meanwhile, the combination therapy can decrease expression of aquaporin-4 and mitigate cerebral edema, which is superior to bone marrow mesenchymal stem cell transplantation and edaravone alone. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Reprogramme-induced genomic stability Cao Ding-ya, Li Jie-liang, Liu Wei-qiang, He Wen-yin, He Wen-zhi, Luo Yu-mei, Fan Yong, Sun Xiao-fang 2014, 18 (10):  1621-1628.  doi: 10.3969/j.issn.2095-4344.2014.10.023 Abstract ( 327 )   PDF (784KB) ( 529 )   Save BACKGROUND: Some studies have shown that more copy number variations are present in early passage human induced pluripotent stem cells than later passage human human induced pluripotent stem cells, their parental somatic fibroblasts or human embryonic stem cells. OBJECTIVE: To investigate whether the reprogramming process itself compromises genomic stability and further explore the efficiency of induced pluripotent stem cell establishment. METHODS: Using high-resolution Affymetrix CytoScan HD array, we compared copy number variations and loss of heterozygosity in early passage induced pluripotent stem cells with their fibroblast cell origins from genetic epilepsy patients. RESULTS AND CONCLUSION: Compared with somatic fibroblasts from genetic epilepsy patient, there was no difference in the loss of heterozygosity between the two types of cells, but more copy number variations were present in early passage human induced pluripotent stem cells which were characterized as microduplication and involved oncogenic genes. Results demonstrate the dynamic nature of genomic abnormalities during reprogramming process and the necessity of frequent monitoring human induced pluripotent stem cells to assure their genomic stability and clinical safety. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Preparation of bone marrow cells-derived extracellular matrix and its microstructure and composition Tang Li, Wu Jun-jie, Wang A-xian, Liang Yuan, Ji Hai-ning, Ding Yin 2014, 18 (10):  1629-1634.  doi: 10.3969/j.issn.2095-4344.2014.10.024 Abstract ( 340 )   PDF (611KB) ( 559 )   Save BACKGROUND: Extracellular matrix can simulate microenvironment and make the stem cells proliferate maintaining the characteristics of stem cells well in vitro.  OBJECTIVE: To prepare the extracellular matrix from human bone marrow cells and to analyze its microstructure and composition preliminarily. METHODS: Human bone marrow cells of passage 4 were cultured for 14 days, and the induction medium was used during the last 8 days. After decellularization, cells were removed to prepare human bone marrow cells-derived extracellular matrix. The surface morphology of human bone marrow cells-derived extracellular matrix was observed by inverted microscope and scanning electron microscope. Changes of collagen I and biglycan before and after decellularization were observed by immunofluorescence staining. Human periodontal ligament stem cells were seeded onto human bone marrow cells-derived extracellular matrix, fibronectin coated 6-well plate and normal culture plate to compare the influence of different matrix on cell morphology and adhesion. RESULTS AND CONCLUSION: We obtained intact human bone marrow cells-derived extracellular matrix by chemical combined with physical decellularization. The structure and amount of collagen I and biglycan had not been compromised dramatically after decellularization. Human periodontal ligament stem cells growing on the  human bone marrow cells-derived extracellular matrix developed in accordance with the orbit of the extracellular matrix, differing from the original cell morphology. There were more human periodontal ligament stem cells adhering to the extracellular matrix during the same time. These findings indicate that effective decellularization can produce intact the extracellular matrix membrane without destroying its microstructure. Extracellular matrix protein is not compromised due to decellularization. The extracellular matrix affects cell morphology and promotes cell adhesion. We can use the extracellular matrix model to simulate stem cell microenvironment and thereafter, acquire a large number of adult stem cells with high quality in vitro. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics

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Stem cell transplantation for diabetic nephropathy: possibility, feasibility and application He Yan-qing, Yang Ping 2014, 18 (10):  1635-1640.  doi: 10.3969/j.issn.2095-4344.2014.10.025 Abstract ( 344 )   PDF (349KB) ( 458 )   Save BACKGROUND: To control blood glucose, blood pressure, blood lipids and inhibit the rennin-angiotensin system is the main idea focused on the treatment of diabetic nephropathy, but the curative effect is unsatisfactory. Hemodialysis and kidney transplantation are suitable for serious cases, however, which is restricted because of the limited source of kidneys and high cost. Regenerative medicine research based on stem cells brings a new hope for treatment of diabetic nephropathy. OBJECTIVE: To comprehensively analyze the mechanism underlying different sources of stem cells for treatment of diabetic nephropathy and the clinical implications. METHODS: Papers addressing stem cells for the treatment of diabetic nephropathy were retrieved by computer in CNKI database and PubMed database from January 2005 to August 2013 with the key words “embryonic stem cells, bone marrow mesenchymal stem cells, induced pluripotent stem cells, diabetic nephropathy" in Chinese and English. Papers published recently or in journals with high impact factor were selected. A total of 60 papers were included for review. RESULTS AND CONCLUSION: Embryonic stem cells, bone marrow mesenchymal stem cells, induced pluripotent stem cells have the potential to differentiate into renal histiocytes. A large numbers of experimental studies have shown that stem cells transplantation has a positive effect on recovery of injured kidney. Stem cell transplantation can provide a novel therapy for diabetic nephropathy. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 全文链接： Figures and Tables | References | Related Articles | Metrics