Characterization and Establishment of an Immortalized Rabbit Melanocyte Cell Line Using the SV40 Large T Antigen

文献类型： 外文期刊

第一作者： Chen, Yang

作者： Chen, Yang;Hu, Shuaishuai;Wang, Manman;Zhao, Bohao;Yang, Naisu;Li, Jiali;Chen, Qiuran;Liu, Ming;Zhou, Juan;Wu, Xinsheng;Chen, Yang;Wu, Xinsheng;Bao, Guolian

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关键词： melanocytes; immortalization; SV40-LT; cell lines; malignant transformation

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

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年卷期： 2019 年 20 卷 19 期

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收录情况： SCI

摘要： Melanocytes (MCs) are specialized cells that synthesize melanin within the melanosome. Cultured MCs are useful in order to study their role in relation to pigmentation. However, MC isolation is laborious and the obtained cells have a limited culture time. In this study, we transformed lentivirus-mediated simian virus 40 Large T (SV40-LT) into primary rabbit melanocytes (Pri RMCs) to establish an immortalized cell line. Morphologically, the immortalized RMCs (Im RMC) were indistinguishable from the Pri RMCs, and dendrites were visible following Dopa staining. No significant differences in cell proliferation or growth between immortalized and primary RMCs were observed. Based on melanocyte-specific markers, the expression of MITF, TYR, and TYRP1 were detected by PCR, immunofluorescence staining, and western blot analysis. Through karyotype, soft agar, and tumorigenesis assays, the immortalized RMCs did not undergo malignant transformation. Our results show that Im RMCs can be used as a tool cell for future MC studies on the pigmentation mechanisms of fur animals.

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