体外诱导脐血间充质干细胞向胰岛β样细胞分化的初步研究

Preliminary Study on the Induction of Umbilical Cord Blood-derived Mesenchymal Stem Cells into Pancreatic Islet β-like Cells in Vitro

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。

Mesenchymal stem cells derived from umbilical cord blood were induced with epidermal growth factor （EGF）, β-mercaptoethanol, high concentration of glucose, activin A and hepatocyte growth factor （HGF）. Insulin in the treated cells was examined by immunofluorescence. In addition, the quantity of insulin secretion and glucose-stimulated insulin release were examined by chemiluminescence immunoassay. Results demonstrated that most of the cells exhibited the change of morphology and produced islet-like cell clusters. Immunofluorescence assay showed insulin positive expression in the induced cells. In addition, chemiluminescence immunoassay demonstrated that induced cells had the reaction to the stimulation of high glucose. Therefore MSCs derived from umbilical cord blood have the potential to be differentiated into islet β-like cells in vitro.