大鼠胎盘来源间充质干细胞的生物学特性

发布时间：2018-09-19 06:20

【摘要】：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。
[Abstract]:Background: it is reported that the results of human mesenchymal stem cells used in experimental animals are not satisfactory, so we consider using mesenchymal stem cells from rat placenta to obtain better results. There are no reports of rat placenta derived mesenchymal stem cells. Aim: to establish a method for isolating mesenchymal stem cells from rat placenta and observe its basic biological characteristics. Methods: mesenchymal stem cells were isolated from rat placenta by collagenase digestion. The shape was observed by inverted microscope every day. The growth kinetics was measured by MTT method and the growth curve was drawn. The phenotype and cell cycle were detected by flow cytometry. The potential of adipogenic and osteogenic differentiation was confirmed by immunohistochemical method. Results and conclusion: after 8 hours of primary cell culture, the cells were colonized within 24 hours. The size and morphology of the fourth passage cells were basically the same, mainly fusiform. Cell cycle detection showed that the proportion of G 2 phase in S phase of G0/G1 phase was 83.768.01 and 8.23%, respectively. Immunophenotypic analysis: expression of CD29 and CD90, does not express CD45. In vitro induction experiments confirmed that placental mesenchymal stem cells had the potential of adipogenic and osteogenic differentiation.
【作者单位】：暨南大学医学院;中国医学科学院血液学研究所泰达生命科学技术研究中心;中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室;广州汉氏联合生物科技有限公司;
【基金】：广东省科技厅省部产学研结合示范基地项目(广东省干细胞产业化基地,项目编号:2010B091101004)支持~~
【分类号】：R329

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