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

Abstract：

: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.

journal_name

Int J Mol Sci

journal_title

International journal of molecular sciences

authors

Chen Y,Hu S,Wang M,Zhao B,Yang N,Li J,Chen Q,Liu M,Zhou J,Bao G,Wu X

doi

10.3390/ijms20194874

subject

Has Abstract

pub_date

2019-09-30 00:00:00

issue

19

issn

1422-0067

pii

ijms20194874

journal_volume

20

pub_type

杂志文章

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