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Establishment and evaluation of a stable steroidogenic goat Leydig cell line
Author(s) -
Zhou Jinhua,
Dai Rui,
Lei Lanjie,
Lin Pengfei,
Lu Xiaolong,
Wang Xiangguo,
Tang Keqiong,
Wang Aihua,
Jin Yaping
Publication year - 2016
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1111/asj.12454
Subject(s) - telomerase reverse transcriptase , leydig cell , telomerase , cholesterol side chain cleavage enzyme , transfection , immortalised cell line , biology , androgen receptor , steroidogenic acute regulatory protein , cell culture , cytochrome p450 , endocrinology , microbiology and biotechnology , medicine , gene , luteinizing hormone , gene expression , genetics , hormone , metabolism , prostate cancer , cancer
Leydig cells play a key role in synthesizing androgen and regulating spermatogenesis. The dysfunction of Leydig cells may lead to various male diseases. Although primary Leydig cell cultures have been used, their finite lifespan hinders the assessment of long‐term effects. In the present study, primary goat Leydig cells (GLCs) were immortalized via the transfection of a plasmid containing the human telomerase reverse transcriptase ( hTERT ) gene. The expressions of hTERT and telomerase activity were evaluated in transduced GLCs (hTERT‐GLCs). These cells steadily expressed the hTERT gene and exhibited longer telomere lengths at passage 55 that were similar to those of HeLa cells. The hTERT‐GLCs at passages 30 and 50 expressed genes that encoded key proteins, enzymes and receptors that are inherent to normal Leydig cells, for example, steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side‐chain cleavage enzyme (P450scc), 3β‐hydroxysteroid dehydrogenase (3β‐HSD) and LH‐receptor (LH‐R). Additionally, the immortalized goat Leydig cells secreted detectable quantities of testosterone in response to hCG stimulation. Furthermore, this cell line appeared to proliferate more quickly than the control cells, although no neoplastic transformation occurred in vitro . We concluded that the GLCs immortalized with hTERT retained their original characteristics and might provide a useful model for the study of Leydig cell function.