Establishment and characterization of dairy cow growth hormone secreting anterior pituitary cell model.
A dairy cow anterior pituitary cell (DCAPC) model was established in vitro for the study of growth hormone (GH) synthesis and secretion in the anterior pituitary gland of the dairy cow. Pituitary glands were obtained from Holstein dairy cows' heads cut by electric saw, and the posterior pituitary glands were removed to obtain integrated anterior pituitary glands. Immunohistochemistry assay of GH in the anterior pituitary glands showed that most somatotrophs were located within the lateral wings of the anterior pituitary. Tissues of the lateral wings of the anterior pituitary were dispersed and cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. The DCAPCs displayed a monolayer, cobblestone, epithelial-like morphology which are the typical characteristics of the anterior pituitary cells. The DCAPCs were subcultured continuously over ten passages. GH immunoreactivity was present in DCAPCs at passage 10. The transcription of the bovine GH mRNA in DCAPCs at passage 10 was decreased to below 50% compared with the lateral wings of the anterior pituitary tissues. Thus, our DCAPCs model is effective for the in vitro examination of GH synthesis and secretion in the dairy cow anterior pituitary gland. The effects of transforming growth factor beta 1 (TGF-β1) and interferon-γ (IFN-γ) on the expression of GH mRNA in DCAPCs at passage 3 were also investigated. There were no obvious changes in transcription of the GH gene after treatment with TGF-β1 for 24 h, while IFN-γ increased transcription of the GH gene in a dose-dependent manner.
College of Veterinary Medicine, Jilin University, Changchun, 130062, People's Republic of China.
Jian-Fa
JF
Wang
Shou-Peng
SP
Fu
Su-Nan
SN
Li
Zhan-Qing
ZQ
Yang
Wen-Jing
WJ
Xue
Zhi-Qiang
ZQ
Li
Wei
W
Wang
Ju-Xiong
JX
Liu
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D005260:Female; D013006:Growth Hormone / Q000096:biosynthesis* / Q000235:genetics; D007371:Interferon-gamma / Q000235:genetics; D010903:Pituitary Gland, Anterior / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000235:genetics
D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; C073294:growth hormone, bovine; D007371:Interferon-gamma; D013006:Growth Hormone