传代后视网膜色素上皮细胞MERTK基因表达变化的研究

摘要

目的:检测传代后的视网膜色素上皮细胞(retinal pigment epithelial,RPE)MERTK基因表达的变化,明确传代的异体RPE移植的可行性.方法:离体培养原代的人视网膜色素上皮细胞,将原代细胞以1:3比例进行传代至第6代,以SYBR(R) Green I real time RT-PCR方法检测不同传代数的RPE 细胞MERTK基因表达的水平,采用2-ΔΔCT 方法计算定量PCR的数值进而比较各代MERTK基因表达的的变化.结果:原代及传代的RPE细胞均表达MERTK基因.第二代RPE细胞MERTK基因表达水平与原代相同,第3~6代RPE细胞MERTK基因表达水平均明显低于原代.结论:传代的RPE细胞可以作为异体视网膜色素上皮细胞移植的细胞来源,但以第二代的RPE细胞为最佳选择.%AIM: To exam the alteration of MERTK gene in different passages and clarify the feasibility of passaged allograft retinal igment epithelial (RPE) cells transplantation.METHODS:Human primary RPE cells were passaged as ratio 1 ∶ 3, the level of MERTK gene in different passages cells were quantified by SYBR Green I real time RT-PCR. The 2-ΔΔCT method was used to calculate relative changes in gene expression determined from real-time uantitative PCR experiments.RESULTS: Both primary and passaged RPE cells expressed MERTK gene. The level of MERTK gene in second passage RPE cells were the same as the primary cells,while in passage 3-6 the level of MERTK gene were declined obviously.CONCLUSION: The passaged allograft RPE cells could be a source for transplantation, but the second passage RPE cells were the etter selected object.