|Chitko Mckown, Carol|
|Mckown, Richard - UNIVERSITY OF NEBRASKA|
Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: September 10, 2004
Publication Date: November 1, 2004
Citation: Chitko Mckown, C.G., Green, B.T., Miller, L.C., Mckown, R.D., Laegreid, W.W. 2004. Development and characterization of three porcine monocyte-derived cell lines [abstract]. Research Workers in Animal Diseases Conference. p. 96. Technical Abstract: The study of macrophage-tropic pathogens is made more difficult in food animals by the paucity of monocyte/macrophage cell lines available from these species. Therefore, it was our objective to develop macrophage-like cell lines from porcine peripheral blood mononuclear cells. Whole blood samples were collected from a cross-bred grower pig into syringes containing EDTA as the anticoagulent. Peripheral blood mononuclear cells were purified using density gradient centrifugation, and monocytes were obtained by removing non-adherent cells from the tissue culture flasks. The cells were divided into three initial treatment groups: no treatment (control monocytes), treatment with L929 supernatant, and treatment with human recombinant macrophage colony stimulating factor. After culturing under these conditions for 17 days, all cell lines were then cultured in tissue culture medium containing from 5 to 20% L929 supernatant. The cells have been grown continuously for greater than three months. Preliminary characterization has shown that these cell lines morphologically resemble macrophages, and positively stain for the production of alpha-naphthyl esterase and negatively stain for peroxidase, consistent with macrophages. Ongoing characterization of these cell lines for cell-surface markers, macrophage activity such as production of inducible nitric oxide synthase, phagocytosis, and bactericidal activity, as well as susceptibility to pathogens will be described.