传代后视网膜色素上皮细胞MERTK基因表达变化的研究(英文)

The alteration of MERTK gene in different passage of retinal pigment epithelium cells

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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 pigment 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-△△CT method was used to calculate relative changes in gene expression determined from real-time quantitative 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 better selected object.