脐带血血清培养人脐带血间充质干细胞  

Cultivation of human umbilical cord blood mesenchymal stem cells with cord blood serum

作　　者：桑宵 吕玲 于丽丽 邢义高 张秀涛 雒猛 SANG Xiao;LYU Ling;YU Lili(Cord Blood Hematopoietic Stem Cell Bank of Shandong Province,Jinan 250102,CHINA)

机构地区：[1]山东省脐带血造血干细胞库,山东济南250102

出　　处：《江苏医药》2023年第10期977-981,共5页Jiangsu Medical Journal

摘　　要：目的探讨利用脐带血血浆分离的脐带血血清(CBS)培养人脐带血间充质干细胞(UCB-MSCs)的可行性。方法从脐带血血浆分离制备CBS,检测其质量参数。培养2例UCB-MSCs,设置3种培养体系,分为五组,分别采用同体积CBS(CBS-1组、CBS-2组、CBS-3组)、FBS(FBS组)和Elite血清替代物(Elite血替组)培养UCB-MSCs至P8代,显微镜下观察UCB-MSCs形态,绘制细胞增殖倍数曲线。采用流式细胞术检测UCB-MSCs表面标记物。结果成功建立脐带血血浆分离CBS的方法,并完成CBS质量参数检测及性能测试。在3种培养体系中UCB-MSCs增殖倍数存在差异,在CBS培养体系中表现出良好的一致性。UCB-MSCs在3种培养体系中均高表达CD90、CD73、CD105,而低表达CD34、CD45及HLA-DR。结论CBS稳定性良好,可代替FBS用于UCB-MSCs的扩增培养。扩增后的UCB-MSCs形态良好,一致性高,可稳定表达MSCs表型。Objective To explore the feasibility of culturing human umbilical cord blood mesenchymal stem cells(UCB-MSCs)with cord blood serum(CBS)isolated from cord blood plasma.Methods CBS was isolated and prepared from cord blood plasma,and its qualities were evaluated.UCB-MSCs were divided into five groups cultured with the same volume of CBS(group CBS-1,group CBS-2and group CBS-3),FBS(group FBS),and Elite serum substitute(group Elite),respectively.The morphology of UCB-MSCs was observed by microscope,and the cell proliferation multiple curve was drawn.Flow cytometry was used to detect the surface markers of UCB-MSCs.Results The method of isolating CBS from cord blood plasma was established successfully,and the quality parameter detection and performance test of the prepared CBS were completed.There were differences in UCB-MSCs proliferation multiplication among the three culture systems.UCB-MSCs exhibited good consistency in the CBS culture system.UCB-MSCs highly expressed CD90,CD73and CD105in different culture systems,while CD34,CD45and HLA-DR were poorly expressed.Conclusion CBS has stable quality and can be used in the amplification culture of UCB-MSCs instead of FBS.The amplified UCB-MSCs exhibit normal morphology and high consistency,and the cell phenotype can be stably expressed.

关 键 词：脐带血血清 脐带血间充质干细胞 细胞培养 

分 类 号：Q813[生物学—生物工程]