Generation and characterization of a microglial cell line, MG5, derived from a p53-deficient mouse.
We have established a cell line cloned from primary-cultured microglia obtained from p53-deficient mice. The cell line, MG5, could be grown in astrocyte-conditioned medium and has been maintained for more than a year. MG5 cells are immunocytochemically positive for Mac-1 and F4/80 antibody and express the major histocompatibility complex (MHC) class I antigen, leukocyte function-associated antigen-1, leukocyte common antigen, and intercellular adhesion molecular-1 mRNA. Interferon-gamma enhanced the expression of MHC class II antigen mRNA in MG5 cells. We previously identified a novel calcium-binding protein, Iba1 (ionized calcium-binding adapter molecule 1), which is highly and specifically expressed in cultured microglia. Iba1 protein was also immunocytochemically demonstrated in MG5 cells. The cells retained non-specific esterase activity, 5'-nucleotidase activity, acid phosphatase activity, and phagocytic ability. Like primary cultured microglia from wild-type mice, MG5 cells released nitric oxide in response to lipopolysaccharide, and actively proliferated in the presence of mitogenic factors such as macrophage-colony stimulating factor (M-CSF), granulocyte/macrophage-CSF (GM-CSF), and interleukin-3 (IL-3). Tyrosine-phosphorylation of M-CSF receptor in MG5 cells was induced by the addition of M-CSF or astrocyte-conditioned medium. These findings indicate that MG5 cells preserve the morphological, biochemical, and physiological properties of primary-cultured microglia well. The MG5 cell line will be a useful tool for studying microglial function.
Ohsawa
K
K
Department of Neurochemistry, National Institute of Neuroscience, Kodaira, Tokyo, Japan.
Imai
Y
Y
Nakajima
K
K
Kohsaka
S
S
D015720:5'-Nucleotidase / Q000096:biosynthesis; D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D004790:Enzyme Induction; D004950:Esterases / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D017628:Microglia / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010641:Phenotype; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency*
D008070:Lipopolysaccharides; D016159:Tumor Suppressor Protein p53; D009569:Nitric Oxide; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D016186:Receptor, Macrophage Colony-Stimulating Factor; D004950:Esterases; D000135:Acid Phosphatase; D015720:5'-Nucleotidase