论文部分内容阅读
背景:研究报道,人间充质干细胞用于实验动物研究的结果不令人满意,所以考虑使用大鼠胎盘来源的间充质干细胞用于动物实验研究,以期获得更好的实验结果。查阅国内外相关文献,目前尚未发现大鼠胎盘来源间充质干细胞的相关报道。目的:建立从大鼠胎盘分离间充质干细胞的方法,观察其基本生物学特性。方法:通过胶原酶消化法从大鼠胎盘分离并得到间充质干细胞。每天用倒置显微镜观察其形态。用MTT法测定其生长动力学并绘制生长曲线。用流式细胞仪检测其表型及细胞周期。通过免疫组化法证实其成脂及成骨分化的潜能。结果与结论:原代细胞培养8h后细胞贴壁,24h内细胞形成集落,第4代细胞大小、形态基本一致,以梭形为主。细胞周期检测提示G0/G1期、S期、G2期的比例分别为83.76%,8.01%,8.23%。免疫表型分析其表达CD29和CD90,不表达CD45。体外诱导实验证实胎盘间充质干细胞具有成脂和成骨分化的潜能。
BACKGROUND: The research reported that the results of using human mesenchymal stem cells in experimental animal research are unsatisfactory. Therefore, the use of rat placenta-derived mesenchymal stem cells for animal experimental research is considered in order to obtain better experimental results. Access to relevant literature at home and abroad, there have been no reports of placental derived mesenchymal stem cells in rats. OBJECTIVE: To establish a method for the isolation of mesenchymal stem cells from rat placenta and to observe their basic biological characteristics. Methods: Mesenchymal stem cells were isolated from rat placenta by collagenase digestion. Morphology was observed daily with an inverted microscope. The growth kinetics was measured by MTT method and the growth curve was drawn. Flow cytometry was used to detect the phenotype and cell cycle. The potential of adipogenic and osteogenic differentiation was confirmed by immunohistochemistry. RESULTS AND CONCLUSION: After cultured for 8 hours, the primary cells adherent to the cells and the cells formed colonies within 24 hours. The size and morphology of the 4th generation cells were basically the same with a spindle shape. Cell cycle test showed that the proportion of G0 / G1 phase, S phase and G2 phase were 83.76%, 8.01% and 8.23% respectively. Immunophenotype analysis of its expression CD29 and CD90, does not express CD45. In vitro induction experiments confirmed that placental mesenchymal stem cells have adipogenic and osteogenic differentiation potential.