大鼠胰腺导管来源干细胞向胰岛素分泌细胞分化的体外培养

Differentiation of rat pancreatic duct-derived stem cells into insulin-secreting cells in vitro

目的:研究在体外条件下从大鼠胰腺导管分离的干细胞向胰岛素分泌细胞分化的产物细胞的形态、表型及功能.方法:采用胶原酶原位消化法消化大鼠胰腺,差异贴壁法培养出胰腺导管来源干细胞(PDSCs),对其进行形态学与表型鉴定.采用无血清培养基,添加Matrigel、exendin-4诱导干细胞向胰岛素分泌细胞分化,鉴定产物细胞的形态、表型及功能.结果:PDSCs在塑料培养皿内贴壁生长,在含血清的培养基内高度增殖,形态学上呈索形或纺锤形的成纤维样细胞,表型上是一种表达PDX-1和nestin的间充质干细胞;经无血清诱导培养,其可分化为胰岛样细胞团并释放胰岛素,但其胰岛素释放指数(SI)为1.35±0.27,明显弱于新鲜分离的胰岛,尚难满足胰岛移植的要求.血清可以促进PDSCs的增殖但抑制其分化,Matrigel与exendin-4对其分化具有促进作用.结论:PDSCs是一种表达PDX-1和nestin的间充质干细胞,可在体外向胰岛素分泌细胞分化,是一种潜在的新的胰岛来源.

AIM：To isolate and identify rat pancreatic ductderived stem cells（PDSCs）and to induce their differentiation into insulin-secreting cells in vitro.METHODS：The pancreas of rats underwent in situ collagenase digestion,and PDSCs were obtained by adherent cultivation.The morphology and phenotypes of PDSCs were characterized.PDSCs were then cultured in serum-free medi-um containing Matrigel and exendin-4 to induce their differentiation into insulin-secreting cells.The morphology,phenotypes,and functions of the obtained insulin-secreting cells were then characterized.RESULTS：Rat PDSCs had a typical fibroblast-like shape,grew rapidly in an adherent manner in serum-containing medium,and belonged to mesenchymal stem cells（MSCs）which expressed PDX-1 and nestin.Serum-free cultivation could induce PDSCs to differentiate into isletlike cell clusters（ICCs）that possessed the capability of secreting insulin.CONCLUSION：Rat PDSCs belong to MSCs which express PDX-1 and nestin and could differentiate into insulin-secreting cells in vitro.