Y27632体外诱导视网膜色素上皮细胞转分化为神经元样细胞的初步研究  

Y27632 induce trans-differentiation from retinal pigment epithelial cells into neuron-like cells in vitro

作　　者：金怡轩[1] 张少冲[1] 郑健樑[1] 董志章[1] 黄雄高[1] 

机构地区：[1]中山大学中山眼科中心眼科学国家重点实验室,广州510060

出　　处：《中华眼底病杂志》2011年第3期275-277,共3页Chinese Journal of Ocular Fundus Diseases

基　　金：广东省科技计划项目（20088030301306）

摘　　要：目的探讨Y27632在体外诱导成人视网膜色素上皮（RPE）细胞转分化为神经元样细胞的可行性。方法取生长良好的第3～6代成人RPE接种于6孔培养板，细胞贴壁生长后分为对照组和实验组。对照组培养基为2％胎牛血清（FBS）＋Dulbeeco改良Eagle培养基（DMEM）／F12培养液，实验组在此培养基中加入10μmol／LY27632。于诱导3、6h和1、3、5、7d观察细胞形态，并用细胞免疫荧光染色法鉴定诱导后3d对照组和实验组角蛋白CKl8、微管相关蛋白Map2、神经丝蛋白NF200、Pax6的阳性表达率，两组间的比较采用γ。检验。结果实验组经诱导后RPE细胞发出轴突样突起并相互连接成网状，呈现典型的神经元细胞形态，占50．1％。对照组RPE和实验组细胞角蛋白表达阳性率分别为93．97％和43．88％，两组比较差异有统计学意义（X^2=64．763，P〈O．01）；Map2对照组和实验组阳性率分别为4．49％和31．90％，两组比较差异有统计学意义（X^2=23．634，P〈0．01）；NF200对照组阳性表达率为22．37％，但阳性表达细胞形态为类圆形，实验组阳性率表达为57．45％，大量细胞发出轴突样突起，两组比较，差异有统计学意义（X^2=21．261，P〈0．01）；Pax6对照组和实验组细胞阳性率分别为8．33％和65．79％，两组比较，差异有统计学意义（X^2=25．946，P〈0．01）。结论Y27632体外诱导成人RPE细胞出现神经元样细胞，是一种可行的诱导方法。Objective To investigate the feasibility of Y27632 to induce transdifferentiation from human retinal pigment epithelial （hRPE） cells into neuron-like cells in vitro. Methods The third to sixth generation of primary hRPE cells were cultured with 2% fetal bovine serum ＋ Dulbecco＇s modified eagle medium/F12 culture solution, with （experimental group） or without （control group） 10 μmol/L Y27632. At 3, 6 hours and 1, 3, 5, 7 days after induction, the morphologic changes of RPE cells were observed by inverted microscope. The expression rate of CK18, Map2, NF200 and Pax6 at 3 days after induction in the experimental and control group were detected by immunofluorescent staining. X^2 test was employed for comparison between the two groups. Results 50. 1 % cells of the experimental group formed axon-like processes and interconnected each other with typical neuron-like appearance. The expression rates of CK18, Map2, NF200 and Pax6 in the experimental group were 43.88%, 31.90%, 57.45% and 65.79%, while the above indexes in the control group were 93.97%, 4.49%, 22. 37% and 8.33% respectively. Compared the expression rate of CK18 （X^2=64. 763）, Map2 （ X^2=23. 634）, NF200 （X^2=21. 261） and Pax6 （X^2= 25. 946） between the two groups, the differences were significant （P〈0.01）. Conclusion The hRPE cells can be trans-differentiated into neuron-like cells in vitro by Y27632.

关 键 词：色素上皮 眼/细胞学 神经元 细胞分化 

分 类 号：R774.1[医药卫生—眼科]