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探讨脐血间充质干细胞能否在体外向胰岛β样细胞分化,并探索其诱导条件。在无菌条件下采集正常产妇脐血,用羟乙基淀粉沉淀法分离脐血中的有核细胞,进而采用贴壁筛选法获得脐血间充质干细胞。纯化后的脐血间充质干细胞用表皮生长因子、β-巯基乙醇、高糖、激活素A和肝细胞生长因子进行诱导。观察诱导后的细胞形态变化,采用胰岛素免疫荧光染色对诱导后的细胞进行鉴定,定量检测胰岛素分泌水平及其对葡萄糖刺激的反应性。结果发现,经过诱导后,细胞形态发生明显变化,形态变圆而且聚集成团;诱导后细胞的胰岛素免疫荧光染色为阳性;而且细胞能分泌少量胰岛素,并对糖刺激具有反应性。由此提示,在体外,脐血间充质干细胞具有向胰岛β样细胞分化的潜能。
To investigate whether umbilical cord blood mesenchymal stem cells can differentiate into pancreatic β-like cells in vitro and to explore the induction conditions. Normal maternal umbilical cord blood was collected under aseptic conditions, and the nucleated cells in umbilical cord blood were separated by hydroxyethyl starch precipitation, and then the umbilical cord blood mesenchymal stem cells were obtained by adherence screening. Purified cord blood MSCs were induced with epidermal growth factor, β-mercaptoethanol, high glucose, activin A and hepatocyte growth factor. The morphological changes of the cells after the induction were observed. Insulin immunofluorescence staining was used to identify the induced cells, and the level of insulin secretion and the reactivity to glucose stimulation were quantitatively determined. The results showed that, after induction, cell morphology changed significantly, the morphology of rounded and clustered into groups; induced by immunofluorescence staining of insulin-positive cells; and the cells can secrete a small amount of insulin, and sugar-responsive. Thus, in vitro, umbilical cord blood mesenchymal stem cells have the potential to differentiate into pancreatic β-like cells.