Detection of interleukin-1 and interleukin-6 on cryopreserved bovine mammary epithelial cells in vitro

J Vet Med Sci. 1997 Jul;59(7):503-7. doi: 10.1292/jvms.59.503.

Abstract

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)-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, interferon (INF)-tau, TNF-alpha, 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-1alpha, IL-1beta, IL-6, IL-10, TNF-alpha, 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cell Division
  • Cells, Cultured
  • Cryopreservation*
  • Epithelial Cells
  • Epithelium / immunology
  • Female
  • Interleukin-1 / analysis
  • Interleukin-1 / biosynthesis*
  • Interleukin-6 / analysis
  • Interleukin-6 / biosynthesis*
  • Kinetics
  • Lactation
  • Mammary Glands, Animal / cytology*
  • Mammary Glands, Animal / immunology*
  • Time Factors

Substances

  • Interleukin-1
  • Interleukin-6