title: Establishment and evaluation of a stable steroidogenic goat Leydig cell line.
abstract: 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.
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Dai
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Lanjie
authors.initials: L
authors.lastname: Lei
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Lin
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Lu
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Xiangguo
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Keqiong
authors.initials: K
authors.lastname: Tang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Aihua
authors.initials: A
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shanxi, China.|College of Veterinary Medicine, Northwest A&F University, Yangling, Shanxi, China.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Jin
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000502:physiology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D006041:Goats; D007985:Leydig Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism / Q000502:physiology; D008297:Male; D010750:Phosphoproteins / Q000378:metabolism; D011974:Receptors, LH / Q000378:metabolism; D013091:Spermatogenesis; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D059505:Telomere Homeostasis; D013739:Testosterone / Q000378:metabolism*; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D006063:Chorionic Gonadotropin; D010750:Phosphoproteins; D011974:Receptors, LH; D000098682:Steroidogenic Acute Regulatory Protein; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase; D002786:Cholesterol Side-Chain Cleavage Enzyme; C509186:TERT protein, human; D019098:Telomerase
keywords: Leydig cell; goat; hTERT; immortalization