传代后视网膜色素上皮细胞MERTK基因表达变化的研究
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国家自然科学基金资助项目(No.39800167)


The alteration of MERTK gene in different passage of retinal pigment epithelium cells
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National Natural Science Foundation of China(No.39800167)

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    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。

    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.

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孙昱昭,彭荣梅,刘经纬,等.传代后视网膜色素上皮细胞MERTK基因表达变化的研究.国际眼科杂志, 2013,13(3):421-424.

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  • 收稿日期:2012-10-18
  • 最后修改日期:2013-02-10
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