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背景:人羊膜间充质干细胞可诱导分化为神经样细胞,但在培养过程中发现干细胞增殖的数量不足。目的:为了获得足够数量的人羊膜间充质干细胞,观察人羊膜间充质干细胞混合培养的生长情况及经碱性成纤维细胞生长因子诱导人羊膜间充质干细胞向神经样细胞的分化情况。方法:将两个不同胎盘来源的羊膜分离培养、混合,采用细胞分离和培养技术获取人羊膜间充质干细胞,分离人羊膜间充质干细胞后传代培养,加入碱性成纤维细胞生长因子诱导分化。当细胞传至第3代时,随机取培养的6份人羊膜间充质干细胞(半量)为A组,6份人羊膜间充质干细胞(半量)为B组,将A、B组各剩余的半量人羊膜间充质干细胞混合为C组。3组细胞浓度均为1.0×107L-1。以活细胞计数和MTT比色法比较3组细胞扩增数量,以免疫组织化学法检测羊膜间充质干细胞神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白的表达。结果与结论:混合人羊膜间充质干细胞之间有互相促增殖的作用。人羊膜间充质干细胞具有较强的可朔性,经碱性成纤维细胞生长因子诱导的人羊膜间充质干细胞可表达神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白。
BACKGROUND: Human amniotic mesenchymal stem cells can induce the differentiation into neuron-like cells, but the number of stem cell proliferation is not enough during the culture. OBJECTIVE: To observe the growth of human amniotic mesenchymal stem cells mixed with human amniotic mesenchymal stem cells and to observe the differentiation of human amniotic mesenchymal stem cells into neuron-like cells induced by basic fibroblast growth factor. Methods: Two amniotic membranes derived from different placenta were isolated and cultured. Human amniotic mesenchymal stem cells (MSCs) were obtained by cell isolation and culture technique. Human amnion mesenchymal stem cells were isolated and subcultured. The addition of basic fibroblast growth factor . When the cells reached the third generation, 6 human amniotic mesenchymal stem cells (half amount) were randomly selected from group A, 6 human amniotic mesenchymal stem cells (group B), and the remaining groups A and B Half of human amniotic mesenchymal stem cells were mixed as group C. Three groups of cells were 1.0 × 107L-1. The numbers of cells in three groups were compared by viable cell count and MTT colorimetric method. The expression of glial cell marker, neuron specific enolase and nestin in amniotic mesenchymal stem cells was detected by immunohistochemistry. RESULTS AND CONCLUSION: The amniotic membrane-derived mesenchymal stem cells have the function of promoting proliferation to each other. Human amniotic mesenchymal stem cells have strong scalability. Human amniotic mesenchymal stem cells induced by basic fibroblast growth factor can express glial cell markers, neuron-specific enolase and nestin.