摘要
目的:比较胶原酶的不同消化时间,观察所获得的间充质干细胞的数量,确定最为理想的消化时间。方法:分别采用3h、5h、7h的消化时间分离脐带间充质干细胞;通过传代进行纯化和扩增培养,绘制生长曲线;用流式细胞仪检测其表面标志。结果:分离出的间充质干细胞贴壁后为梭形,呈平行排列生长或漩涡状生长,并且消化时间为5h时收获到的细胞数量最多;流式细胞仪检测结果显示,获得的间充质干细胞均表达CD73、CD105、HLA-ABC,不表达CD34、CD45、HLA-DR。结论:胶原酶消化法从人脐带中分离培养的细胞具有间充质干细胞的生物学特性,并且消化5h收获得的间充质干细胞数量最多,生长状况最好。
Objective Comparing the quantity of MSCs acquired from umbilical cord that digested by collagenase Ⅱ in different hours,ascertain an ideal digesting time and culture and appraise the cells acquired.Methods MSCs were isolated from human umbilical cord through collagenase digestion respectively in 3 h,5 h,7 h,then passaged to purify and culture,drew the growth curve.Their surface markers and cells cycle were detected by flow cytometry.Results MSCs isolated from umbilical cord ad-hered with a spindle-shape,they parallel along their longitudinal axis or grew in whirl manner.After 5 hours digestion,we got the biggest quantity with best growth status,and successfully built the method for MSCs culture and increasing.Detected by flow cytometry,all of adherent cells express CD73、CD105、HLA-ABC,none of them express CD34、CD45、HLA-DR.Con-clusion Cells isolated from umbilical cord by digestion have the biological characteristics of MSCs.And 5 hours is the best time for digestion to acquire the most and best MSCs.
出处
《中国民族民间医药》
2014年第24期36-40,共5页
Chinese Journal of Ethnomedicine and Ethnopharmacy
关键词
胶原酶
脐带
间充质干细胞
分离
消化时间
Collagenase
Umbilical cord
Mesenchymal Stem Cells
Isolate
Digestion time
