磷脂酰肌醇3激酶抑制剂LY294002诱导人胚胎干细胞定向分化为胰岛素分泌细胞的研究

The study of phosphoinositide-3-kinase inhibitor LY294002 in the differentiation of human embryonic stem cells into more mature insulin-producing cells

目的:探讨磷脂酰肌醇3激酶抑制剂LY294002对人胚胎干细胞定向分化为成熟胰岛素分泌细胞的影响。方法:体外通过5个阶段诱导人胚胎干细胞定向分化为胰岛素分泌细胞。分别给予尼克酰胺+B27(B27组)为对照组和尼克酰胺+LY294002(LY组)为实验组诱导胰岛素分泌细胞的成熟。显微镜下观察各阶段细胞形态变化,免疫荧光染色鉴定胰岛素、c-肽、生长抑素和胰高血糖素的表达。结果:第5阶段诱导14 d LY组胰岛素单染阳性率与B27组无统计学差别(P﹥0.05),但生长抑素和胰高血糖素单染阳性率,胰岛素/生长抑素共染率均低于B27组(P﹤0.05)。结论:在无血清培养体系下,磷脂酰肌醇3激酶抑制剂LY294002能够诱导人胚胎干细胞分化为更加成熟的胰岛素分泌细胞。

Objective To investigate the effect of phosphoinositide-3-kinase inhibitor LY294002 on the differentiation of human embryonic stem cells （HESC） into more mature insulin-producing cells. Methods HESCs were induced to differentiate into insulin-producing cells through five stages. Nicotinamide and B27 （group B27）, nicotinamide and LY294002 （group LY） were used to induce the nesting positive cells into mature insulin-producing cells. The morphological change of each stage was observed under microscope, and expressions of insulin, c-peptide, somatostatin and glucagon were identified by immunofluorescence staining. Results After 14 days in stage 5, there was no significant difference in rate of insulin positive cells between group LY and group B27 （P 〉 0.05）, but rates of somatostatin and glucagon positive cells in group LY were lower than those in group B27（P 〈 0.05）. Furthermore, the co-stained rate of somatostatin and insulin in group LY was also lower than that in group B27 （P 〈 0.05）. Conclusion HESCs can be induced to differentiate into more mature insulin-producing cells by phosphoinositide-3-kinase inhibitor LY294002 in serum-free culture medium.