培养体系对维持人脐带间充质干细胞特性及其体外扩增效率的影响

发布时间：2018-01-18 01:33

本文关键词：培养体系对维持人脐带间充质干细胞特性及其体外扩增效率的影响　出处：《中国组织工程研究》2017年13期 　论文类型：期刊论文

【摘要】：背景:前期数据显示,应用人血小板裂解液培养体系对人脐带间充质干细胞进行体外培养时,较无血清培养体系能更好地维持干细胞特性,而无血清培养体系相较于人血小板裂解液培养体系能一定程度上提高间充质干细胞的体外增殖能力。目的:筛选能最大限度维持人脐带间充质干细胞特性及扩增效率高的培养体系。方法:将6份人脐带标本分别接种于6种培养体系中,进行脐带间充质干细胞原代培养,6种培养体系分别为MesenC ult无血清完全培养基中添加2%人血小板裂解液(A组)、StemP ro无血清完全培养基中添加2%人血小板裂解液(B组)、MesenC ult无血清完全培养基(C组)、Stem Pro无血清完全培养基(D组)、低糖DMEM中添加体积分数10%胎牛血清(E组)、低糖DMEM中添加10%人血小板裂解液(F组),14 d后进行消化传代培养,比较各培养体系对人脐带间充质干细胞形态、表面标记物、分化及增殖能力的影响。结果与结论:(1)D组第3代细胞细长,大小不均;E、F组第3代细胞扁平,其余组第3代细胞为长梭形且大小较均一。A、B组第5代细胞形态及大小较第3代时无明显变化,C组第5代细胞趋于扁平且大小不均一,D组第5代细胞形态较第3代时细长,E组第5代细胞形态较第3代时扁平,F组第5代细胞形态较第3代时无明显变化;(2)各组细胞表面标志物检测结果无明显区别;(3)A、B组成脂、成骨诱导率较高,E、F组次之,C、D组最低;(4)A组各代次细胞扩增数量显著高于C组,B组各代次细胞扩增数量显著高于D组,E、F组扩增数量显著低于培养组A、B组;(5)结果表明,无血清和人血小板裂解液相结合的培养基能较好地维持间充质干细胞的特性及扩增效率。
[Abstract]:Background: previous data showed that human umbilical cord mesenchymal stem cells cultured in vitro with human platelet lysate culture system can better maintain stem cell characteristics than serum-free culture system. Compared with human platelet lysate culture system, serum-free culture system can improve the proliferation of mesenchymal stem cells in vitro. Methods: 6 human umbilical cord specimens were inoculated into 6 culture systems. The primary culture system of umbilical cord mesenchymal stem cells was MesenC ult serum-free complete medium supplemented with 2% human platelet lysate (group A). 2% human platelet lysate group B was added to StemP's serum-free complete medium, and Mesen C ult was added to the complete serum-free medium (group C). Stem Pro serum-free complete medium was added to group D, low glucose DMEM added 10% fetal bovine serum to group E, and low glucose DMEM to 10% human platelet lysate F). The effects of different culture systems on morphology, surface marker, differentiation and proliferation of human umbilical cord mesenchymal stem cells were compared after 14 days. Uneven size; The cells of the third passage were flat in group E and F, and the cells of the third passage in the other groups were fusiform and the size of the cells in the fifth passage of group A B had no significant change compared with that in the third generation. The fifth passage of cells in group C tended to flatten and the size of group D was uneven. The morphology of the fifth passage of group D was more flat than that of group E at the third generation, and that of group E was more flat than that of group E at the third generation. In group F, there was no significant change in the morphology of the fifth passage compared with that in the third generation. 2) there was no significant difference in the detection results of cell surface markers in each group. The ratio of osteogenic induction was higher in group F than that in group C (P < 0.05). The number of expanded cells in group A was significantly higher than that in group C and group B was significantly higher than that in group D and group A was significantly lower than that in group Agna B. The results showed that the liquid phase binding medium without serum and human platelet cleavage could maintain the characteristics and amplification efficiency of mesenchymal stem cells.
【作者单位】：青岛市立医院卫生部细胞移植重点实验室临床中心;
【分类号】：R329.2
【正文快照】： 文章快速阅读:不同培养体系中人脐带间充质干细胞的特性及体外扩增效率消化传代培养结果表明:6组培养体系:原代培养:14 d后:无血清和人血MesenC ult无血清完全培养基中添加2%人血小板裂解液;比较各培养体小板裂解液相脐带间充质StemP ro无血清完全培养基中添加2%人血小板裂解

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