人脐带间充质干细胞分离培养及向脂肪与成骨细胞的分化**△○

背景：研究报道显示人脐带间充质干细胞体外分离方法及效率、培养条件各不相同，并且尚未有统一的鉴定标准。因此建立高效、经济的培养体系十分必要。 目的：本研究旨在建立从人脐带组织分离培养间充质干细胞的方法，并进行向脂肪细胞和成骨细胞的诱导分化。方法：首先采用组织块贴壁法或酶消化法分离纯化得到脐带间充质干细胞(Umbilical cord mesenchymal stem cells,UCMSCs)，分析其细胞形态、增殖方式和某些免疫表型，并在体外诱导其向脂肪细胞和成骨细胞分化。结果和结论:结果发现，hUCMSCs在适当的诱导条件下能向脂肪和成骨细胞分化，体外能连续传代40次以上，且形态和表型保持稳定。这证实了人脐带组织存在间充质干细胞，且具有向脂肪和成骨细胞分化的潜能，该细胞可用于后续的应用研究。

Isolating and differention into adipocytes and osteblasts of human umbilical cord mesenchymal stem cells

BACKGROUND: Culture condition, isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells, which lack of unified identification standards. Therefore, it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem cells (hUCMSCs).OBJECTIVE: To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts. METHODS: The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase. The morphology, proliferation and immunophenotype of the 3rd passage cells were analyzed, and then cells were induced to osteogenic and adipogenic differentiation in vitro.RESULTS AND CONCLUSION: The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro. Flow cytometry analysis revealed that the hUCMSCs were positive for CD29, CD44, CD59, CD105, but were negative for CD40, CD86 and HLA-DR. These cells could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions. The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro. This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.