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  

Effect of cultivation systems on the maintenance of human umbilical cord mesenchymal stem cell characteristics and their proliferation rate in vitro

作　　者：孙亚如[1] 张炳强[1] 王福斌[1] 许评[1] 王二朴[1] 李翠翠[1] 

机构地区：[1]青岛市立医院卫生部细胞移植重点实验室临床中心,山东省青岛市266000

出　　处：《中国组织工程研究》2017年第13期2023-2028,共6页Chinese Journal of Tissue Engineering Research

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。BACKGROUND：Preliminary data showed that the application of human platelet lysate to human umbilical cord mesenchymal stem cell culture can better maintain the characteristics of stem cells than the application of serum-free medium. However, the serum-free medium can better improve the proliferation of mesenchymal stem cells in vitro than the human platelet lysate.OBJECTIVE：To screen out a better mesenchymal stem cell cultivation system that can greatly maintain the characteristics and proliferation rate of human umbilical cord mesenchymal stem cells.METHODS：Six human umbilical cord specimens were inoculated in six culture systems, and the primary culture of umbilical cord mesenchymal stem cell was performed. These six culture systems were respectively MesenCult serum-free medium with 2% human platelet lysate （group A）, StemPro serum-free medium with 2% human platelet lysate （group B）, MesenCult serum-free medium （group C）, StemPro serum-free medium （group D）, low glucose-DMEM with 10% fetal bovine serum （group E）, low glucose-DMEM with 10% human platelet lysate （group F）. The cells were subcultured at 14 days after inoculation to compare the effects of different culture systems on the morphology, surface markers, differentiation and proliferation of human umbilical cord mesenchymal stem cells.RESULTS AND CONCLUSION：（1） The morphology of passage 3 cells in group D was elongated and uneven in size. The morphology of passage 3 cells was flattened in groups E and F, but the cells in the other groups were spindle-shaped and uniform. There were no significant changes in morphology and size between passage 3 and 5 cells in A and B. In group C, the morphology of passage 5 cells was more flattened and uneven in size compared with passage 3 cells. In group D, the morphology of passage 5 cells was more elongated than that of passage 3 cell. In group E, the morphology of passage 5 cells was more flattened than that of passage 3 cells. There was no significant difference in morphology betwe

关 键 词：干细胞 脐带 间质干细胞 组织工程 脐带脐血干细胞 间充质干细胞 培养体系 培养基 筛选 形态 表型 分化 扩增 

分 类 号：R394.2[医药卫生—医学遗传学]