Assay using macrophages for immune response stimulating activity of ingredients derived from chicken eggs
2003
Houkiyama, H. (Hokkaido. Animal Research Center, Shintoku (Japan)) | Noda, Y.
Simple assays for immune response stimulating activity were considered by analyzing proliferation and cytotoxicity of cultured macrophages that were activated by lipopolysaccharide (LPS). The assay was applied to estimate activity of ingredients derived from chicken eggs. J774.1, RAW264, and THP-1 were used for the assays. Cel1 proliferation was evaluated according to increases in the number of cells and morphology. Cytotoxicity was evaluated by generation of nitric oxide. Results showed that J774.1 and RAW264 stopped cell growth and induced morphological differentiation to activated-macrophages. For the cell proliferation assay, it was necessary to culture J774.1 or RAW264 for 3 d and to use 0.4 micro g/ml of LPS or more as a positive control. For the cytotoxicity assay, it was required to culture RAW264 (1 * 10**6 cells/ml) for 24 h and to use 1 micro g /ml of LPS as a positive control. Then, the cytotoxicity assay for immune response stimulating activity was applied to estimate activity of ingredients derived from chicken eggs. It was identified that protease-digested ovomucin, yolk membrane, and chalazae increase generation of nitric oxide. This assay can be used for primary screening of immune response stimulating activity.
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