Sertoli细胞与成人胰岛细胞共同培养的研究

Coculture of Sertoli cells with human islet cells in vitro

目的观察塞尔托利(Sertoli)细胞对体外培养的成人胰岛细胞形态、存活率及功能的影响。方法胰腺、睾丸取自志愿捐赠的成年男性尸体多器官供者,共12例。分离纯化后的成人胰岛细胞分为单独培养组和共同培养组,单独培养组取成人胰岛细胞单独培养,共同培养组为成人胰岛细胞+Sertoli细胞共同培养,均在RPMI1640培养液培养14d,采用倒置相差显微镜观察胰岛细胞形态,比较两组的胰岛细胞存活率、胰岛素分泌量和胰岛素刺激指数。结果培养14d后,共同培养组胰岛细胞存活率为(90±3)%,较单独培养组的(57±4)%明显提高(P<0.01),胰岛细胞的形态亦较单独培养组完整。共同培养组胰岛细胞始终对葡萄糖刺激保持较高的敏感度,而单独培养组胰岛细胞对葡萄糖刺激的敏感度随时间的延长明显降低(P<0.05)。培养14d后,共同培养组的胰岛素分泌量为(249±12)mIU/L、胰岛素刺激指数为8.15±0.64,而单独培养组则分别为(47±7)mIU/L和1.68±0.34,两组比较差异有统计学意义(均为P<0.01)。结论成人胰岛细胞与Sertoli细胞共同培养可以提高胰岛细胞的存活率,改善胰岛细胞的功能。

Objective To investigate the influence of Sertoli cells on the cellular morphology,livabi-lity and function of human islet cells in vitro.Methods Twelve samples of pancreas and testis were taken from the donations of adult males＇ bodies.The purified human islet cells were separated into two groups：control group（human islet cells）and coculture group（human islet cells and Sertoli cells）.The two groups were cultured in culture media of RPMI 1640 for 14 days.The cellular morphology of islet cells was observed by inverted phase contrast microscope.The livability of islet cells,insulin secretory volume and stimulation index was compared between the two groups.Results After cultured for 14 days,the livability of islet cells in coculture group was（90±3）%,while that in control group was（57±4）%（P0.01）.The cellular morphology of islet cells in coculture group was more integrated than that in control group.The sensitivity of islet cells to glucose stimulation was higher in the coculture group,while that was decreased in the control group（P0.05）.After cultured for 14 days,insulin secretory volume and stimulation index were respectively（249±12）mIU/L and 8.15±0.64 in coculture group,while that were respectively（47±7）mIU/L and 1.68±0.34 in control group.There was significant difference between these two groups（all in P0.01）.Conclusion Coculture of human islet cells with Sertoli cells is a promising way to increase livability and improve function of human islet cells in vitro.