协和干细胞基因工程有限公司，天津 300384

    不同消化时间下胶原酶对人脐带间充质干细胞分离的影响

    郝世凯苏相相洪敬欣

    协和干细胞基因工程有限公司，天津 300384

    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。

    胶原酶；脐带；间充质干细胞；分离；消化时间

    Abstract:ObjectiveComparing 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.MethodsMSCs were isolated from human umbilical cord through collagenase digestion respectively in 3h, 5h, 7h, then passaged to purify and culture, drew the growth curve.Their surface markers and cells cycle were detected by flow cytometry.ResultsMSCs isolated from umbilical cord adhered with a spindle-shape, they parallel along their longitudinal axis or grew in whirl manner. After 5hours 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 ......