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.χ2 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(χ2=64.763),Map2(χ2=23.634),NF200(χ2=21.261)and Pax6(χ2=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 bv Y27632. 相似文献
方法:采用Keratograph测量包括首次泪膜破裂时间(the first tear film break-up time, BUT-f)和平均泪膜破裂时间(the average tear film break-up time, BUT-ave)在内的非侵入式泪膜破裂时间(noninvasive tear film break-up time, NI_BUT)结果。测量结果的重复性使用变异系数(coefficient of variation,CV)和组内相关系数(intraclass correlation coefficient,ICC)进行评价, NI-BUT与传统的荧光素泪膜破裂时间(fluorescein tear film break-up time, FBUT)的比较采用Wilcoxon符号秩和检验,确定NI-BUT与FBUT,Schirmer Ⅰ试验结果的相关关系,采用Bland-Altman分析进行一致性评价。
结果:本研究纳入了48个受检者(48眼),平均年龄38.7±15.2岁。BUT-f的CV和ICC分别为12.6%和0.95,BUT-ave的为9.8%和0.96。BUT-f值低于FBUT值,差异有统计学意义(6.16±2.46s vs 7.46±1.92s, P < 0.01)。NI-BUT与FBUT,Schirmer Ⅰ试验结果之间存在显著的正相关关系(P<0.01)。BUT-f与FBUT的95%一致性界限(limits of agreement, LoA)范围为4.46s,BUT-ave与FBUT的LoA范围为3.64s。