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目的从人脐带中获取间充质干细胞(MSC)并进行标记,为进一步移植治疗奠定基础。方法运用组织块贴壁法从人脐带中分离MSC,通过传代培养后,进行形态学观察;采用流式细胞仪对人脐带间充质干细胞(h UC-MSC)的细胞表型进行鉴定;通过特异性染色鉴定体外成脂、成骨诱导后的分化能力;运用免疫荧光证实Brd U标记细胞是否成功。结果运用组织块贴壁法可成功从人脐带中分离出MSC,培养1周左右可见细胞从组织块边缘爬出,向外增殖生长,形成集落;流式细胞仪检测P5代细胞,高表达CD73、CD90及CD105,不表达或低表达CD14、CD34、CD45、CD79a及人类白细胞抗原-DR;成脂诱导2周,油红O染色为阳性,成骨诱导3周,茜素红染色出现红色沉淀物;Brd U标记细胞经免疫荧光检测后,可见红色荧光。结论运用组织块贴壁法从人脐带中分离的细胞,具备MSC的特点并可成功被标记,可用于细胞移植领域的研究。
Objective To obtain and label mesenchymal stem cells (MSCs) from human umbilical cord for the purpose of further transplantation. Methods MSCs were isolated from human umbilical cord by tissue block method. Morphology was observed by subculturing. The cell phenotype of human umbilical cord mesenchymal stem cells (hUC-MSC) was identified by flow cytometry. Specific staining was used to identify adipogenic and osteogenic differentiation in vitro. Immunofluorescence was used to confirm the success of BrdU labeled cells. Results MSC could be successfully isolated from human umbilical cord by tissue block adhesion method. After culturing for about 1 week, the cells climbed out from the edge of the tissue and proliferated and formed colonies. Flow cytometry detected the expression of CD73 , CD90 and CD105 without expression or low expression of CD14, CD34, CD45, CD79a and human leukocyte antigen-DR; positive for oil red O staining for 2 weeks, osteogenic induction for 3 weeks, alizarin red staining for red precipitate ; BrdU labeled cells were detected by immunofluorescence, the red fluorescence. Conclusion The cells isolated from human umbilical cord by tissue block adhesion method have the characteristics of MSC and can be successfully labeled and can be used in the field of cell transplantation.