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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 phosphatidylinositol 3 kinase inhibitor LY294002 on the directional differentiation of human embryonic stem cells into mature insulin-secreting cells. Methods: Human embryonic stem cells were induced to differentiate into insulin-secreting cells through five stages in vitro. Nicotinamide + B27 (B27 group) was given as control group and nicotinamide + LY294002 (LY group) as experimental group to induce the maturation of insulin-secreting cells. The morphological changes of the cells were observed under the microscope, and the expression of insulin, c-peptide, somatostatin and glucagon were identified by immunofluorescence staining. Results: There was no significant difference between the two groups (P> 0.05). However, the positive rates of somatostatin and glucagon staining, insulin / somatostatin staining rate Lower than B27 group (P <0.05). CONCLUSION: Phosphatidylinositol 3 kinase inhibitor LY294002 induces human embryonic stem cells to differentiate into more mature insulin-secreting cells in a serum-free culture system.