人脐带间充质干细胞体外培养的生物学特性研究

目的建立人脐带间充质干细胞(hUC-MSCs)体外培养的方法,探讨该方法培养的脐带间充质干细胞特性及分化潜能。方法用新取的脐带制成细胞悬液,放入含10%胎牛血清的DMEM/F-12混合培养基培养,动态观察原代培养细胞的生长过程。取生长状态良好的第三代细胞(P3),MTT检测细胞增殖情况;流式细胞仪检测表面标志物CD29、CD44、CD31、CD45、CD34,鉴定hUC-MSCs;诱导hUC-MSCs分化成为脂肪细胞,鉴定所分离的hUC-MSCs具有多向分化能力。结果来源于人脐带培养的MSCs呈现梭形生长,经流式细胞仪检测示CD29(98.3%)、CD44(99.2%)阳性,CD34(0.3%)、CD31(0.8%)、CD45(0.4%)阴性,经过诱导分化,证实hUC-MSCs有成脂分化潜能。结论采自脐带标本分离培养可获得稳定、均质性良好的hMSCs,并具有多向分化能力。

Phenotypic and Proliferative Features of Human Umbilical Cord Mesenchymal Stem Cells in Vitro

Objective To establish an in vitro method to culture stem cells derived from human umbilical cord and study the biological characteristics and differentiation potentials of this mesenchymal stem cells.Methods The fresh human umbilical cord was made into cell suspension and cultured in DMEM/F-12 mixed medium which contained l0%fetal bovine serum.The adhesion and proliferation of primary cells were dynamically observed.The P3 hUC-MSCs populations were collected,then the cellular proliferation capacity was detected by thiazolyl blue tetrazolium bromide(MTT) assay.The molecular markers(CD29,CD44,CD31,CD45 and CD34)on the cell surface were identified by flow cytometry(FCM).Oil red O stains assay was to confirm the multilineage potentials of hUC-MSCs.Results The hUC-MSCs isolated from human umbilical cord stromas exhibited fibroblastie morphology and they were positive for CD29(98.3%),CD44(99.2%),and negative for hematopoietic stem cells surface markers CD34(0.3%),CD31(0.8%) and CD45(0.4%).To induce the differentiation of hUC-MSCs toward adiposity was to confirm differentiation potentials of this cultured mesenchymal stem cells.Conclusion The hUC-MSCs are stably isolated and cultured in vitro and confirmed to be differentiated.