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目的探讨脐血来源的人间充质干细胞(HMSCs)在体外分离培养与扩增的可行性。方法在无菌条件下收集正常足月胎儿的脐带血,经复合枸橼酸钠抗凝,用相对密度为1.077g/L的Ficoll淋巴细胞分离液分离脐血的单个核细胞,以30%胎牛血清进行培养和扩增,用流式细胞仪检测MSCs的表面标志。结果来源于人脐血的单个核细胞接种于特定培养基后,可产生贴壁细胞,主要表现为破骨样细胞和间充质样细胞,经传几代后,可得纯化扩增的人脐血MSCs。流式细胞仪检测结果显示,人脐血MSCs不表达CD14、CD19,强表达CD105、CD44。结论来源于人脐血的MSCs在体外可以分离培养、扩增,为MSCs的进一步研究奠定基础。
Objective To investigate the feasibility of isolation, culture and amplification of human umbilical cord blood-derived human mesenchymal stem cells (HMSCs) in vitro. Methods Umbilical cord blood of normal full-term fetuses were collected under aseptic conditions. After anticoagulation with compound sodium citrate, cord blood mononuclear cells were isolated from Ficoll lymphocytes with a relative density of 1.077g / L, Bovine serum was cultured and expanded, and the surface markers of MSCs were detected by flow cytometry. Results The mononuclear cells derived from human umbilical cord blood could produce adherent cells after being inoculated into specific culture medium, mainly expressed as osteoclast-like cells and mesenchymal-like cells. After several passages, purified and expanded human umbilicus Blood MSCs. Flow cytometry results showed that human cord blood MSCs do not express CD14, CD19, strong expression of CD105, CD44. Conclusion MSCs derived from human umbilical cord blood can be isolated, cultured and expanded in vitro, which lays the foundation for further study of MSCs.