Detection of Interleukin-1 and Interleukin-6 on Cryopreserved Bovine Mammary Epithelial Cells in Vitro

Journal of Veterinary Medical Science

Rikio KIRISAWA⁴  Hiromichi OHTSUKA⁵  Takashi ITO¹  Tomo-o YOSHINO²  Hiroshi IWAI⁴  Thomas J. ROSOL³  Hiroyuki OKADA²  Kazuto YAMASHITA⁶ 

[1] Hokkaido Branch, National Institute of Animal Health;Department of Veterinary Pathology, Rakuno Gakuen University;Department of Veterinary Biosciences, The Ohio State University;Department of Veterinary Microbiology, Rakuno Gakuen University;Department of Veterinary Internal Medicine, Rakuno Gakuen University;Department of Veterinary Surgery, Rakuno Gakuen University

关键词: bovine;    cytokine;    mammary;   

DOI  :  10.1292/jvms.59.503

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(24)Cited-By(19)This investigation was performed to determine whether primary cultures of mammary cells from lactating cows would sustain production of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF), and express mRNA for cytokines interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, interferon (INF)-τ, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by the reverse transcription-polymerase chain reaction (RT-PCR) in vitro. Cryopreserved mammary epithelial cells collected from cows at 1 week post calving were plated in collagen-coated 24-well culture plates (250,000 cells/well). IL-1 and IL-6 productions were measured using a A375 cell growth inhibition assay and a 7TD1 hybridoma proliferation assay, respectively. Production of IL-1 was demonstrated in mammary epithelial cells cultured with unsupplemented medium, but was not produced by cells cultured in medium supplemented with fetal bovine serum. IL-6 production in the conditioned medium was continued at steady level until day 14, whereas IL-6-like bioactivity was not detected in medium alone. TNF-like activity was not detectable in any experiments. This study also demonstrated the expression of mRNA for multiple cytokines including IL-1α, IL-1β, IL-6, IL-10, TNF-α, and GM-CSF by RT-PCR in mammary cell cultures. The results indicate that bovine mammary epithelial cells of lactating cows produce IL-1 and IL-6 and have gene expression for multiple cytokines. This in vitro model will be useful to investigate the function and regulation of IL-1 and IL-6 in the lactating mammary gland.

【 授权许可】

Unknown