[关键词]
[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®； 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。
[Key word]
[Abstract]
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-ΔΔCT 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.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(No.39800167)
