目的：探讨人脐带和胎盘来源间充质干细胞（ MSCs）的分离培养和生物学性状的差异，为MSCs的临床应用提供选择依据。方法利用组织贴壁法分离培养脐带MSCs，酶消化法分离培养胎盘MSCs。传代培养后于倒置显微镜下观察两种不同来源的MSCs的细胞形态，流式细胞仪检测两者表面标志物表达的差异，通过细胞周期和群体倍增时间测定比较两者生长增殖能力，体外成骨和成脂诱导比较其分化能力。结果胎盘MSCs和脐带MSCs为贴壁生长、形态均一的成纤维样或长梭形外观，两种细胞均高表达CD90、CD105，不表达CD34、CD45、HLA-DR。胎盘MSCs的群体倍增时间为40．8 h，52．12％处于G0／G1期，脐带MSCs的群体倍增时间为39．5 h，57．50％处于G0／G1期，表明两者增殖能力旺盛，且无明显差异；两者在体外均可诱导分化成骨细胞和脂肪细胞。结论胎盘源MSCs具有与脐带源MSCs相似的生物学特性，两者均可作为临床治疗用细胞或组织工程的种子细胞。%Objective To compare two sources of mesenchymal stem cells ( MSCs) from human placenta and umbilical cord, and to optimize a technical solution for bench or clinical studies of MSCs.Methods MSCs were isolated from human placenta and umbilical cord and expanded for analysis.The cell morphology was observed under invert microscope, the immunophenotypic feature of MSCs was analyzed with flow cytometer, the cell proliferation ability was determined by cell cycle assay and cell doubling time, the cell differentiation potential was evaluated by osteogenic and adipogenic induction in vitro as well.Results Both sources of MSCs were adherent cells and exhibited fusiform and fibrous morphology. Furthermore, both MSCs high expressed CD90 and CD105, and were negative for the markers of CD34, CD45 and HLA-DR.The population doubling time of MSCs form human placenta and umbilical cord was 39.5 h and 40.8 h separately, and the results of cell cycle analysis showed that the percent of the two sources of MSCs in G0/G1 phase was 52.12%and 57.50% respectively. The above results demonstrated that both sources of MSCs possessed the similar biological characteristics in morphology, phenotype and as well as proliferation ability.In addition, both of them could be induced into osteoblasts and adipocytes in vitro.Conclusion MSCs from human placenta have the similar biological characteristics to these from human umbilical cord, and both of them are better candidates for bench and clinical research.
展开▼
