昆明小鼠胰腺干细胞分离培养及特性鉴定

Isolation, culture and identification of pancreatic stem cells of Kunming mouse

目的 分离培养昆明小鼠胰腺干细胞并进行鉴定和观察其分化胰岛细胞的能力。方法 用酶消化法分离生后不同时期昆明小鼠胰腺干细胞 ,用细胞角质蛋白 19(Cytokeratin 19,CK19)、胰岛素 (Insulin)和胰高血糖素 (Glucagon)抗体进行免疫细胞化学染色 ,鉴定细胞种类和分化特性。结果 分离初 ,可见少量上皮样细胞存活 ,更换含有生长因子的培养基后 ,可见细胞分裂增殖 ,呈长梭形 ,逐渐形成集落。传代后鉴定呈CK19阳性。高糖诱导分化后 ,细胞形成胰岛样结构 ,免疫细胞化学染色显示大部分细胞胰岛素阳性 ,少数细胞呈胰高血糖素阳性。结论 昆明小鼠胰腺干细胞能在体外培养条件下增殖 ,具有分化形成胰岛 β细胞和α细胞的能力。

Objective To isolate and identify pancreatic stem cells of Kunming mouse and to observe the differentiation potency of those cells. Methods Pancreas of postnatal Kunming mice were digested by enzymes to isolate pancreatic stem cells. The potency of the cell differentiation was then identified with special marker of cytokeratin 19(CK 19), insulin and glucagons by immunocytochemical staining. Results Few epithelioid cells could be found to survive at the beginning of isolation, but when medium was replaced by that with growth factor, the cells proliferated quickly in fusiform shape and formed colony gradually. The cells were CK 19 immunoreactive positive after transfer of culture. After differentiation induced by high glucose, the cells formed the pancreatic islet like structures. Immunocytochemical staining showed that part of the cells of pancreatic islet like structures were insulin immunoreactive positive and few of the cells were glucagon immunoreactive positive. Conclusion Pancreatic stem cells of Kumming mouse can proliferate when cultured in vitro and have the potency of differentiation into β andα cells of pancreatic islet.