Immortalization and characterization of rat microglial cells.
Microglial cell lines from rat brain were established by transfer of a temperature sensitive simian virus 40 large tumour antigen by means of a retrovirus. Four weeks after infection, colonies were generated in the presence of neomycin and granulocyte-macrophage colony stimulating factor (GM-CSF), and subsequently subcloned. Both bulk cell lines and clones proliferate actively at 33 degrees C, whereas the rate of division was significantly decreased at 39 degrees C when the large T antigen is non-functional. At 39 degrees C, these cells take on the microglial phenotype as demonstrated by immunoreactivity to ED-1 (an intracellular antigen), OX-42 (complement type 3 receptor), W3/25 (CD4 homologue), OX-6 (MHC class II antigen) and OX-18 (MHC class I antigen). These cells are capable of active phagocytosis and retain these properties for 10-15 passages. Long-term culture of these lines and clones, greater than 15 passages, displayed a gradual down-regulation of all cell surface specific antigens that were not rescued by lipopolysaccharide (LPS), interferon-gamma (gamma-IFN), GM-CSF or colony-stimulating factor-1 (CSF-1). The expression of the SV-40 large T antigen was unaffected. These results demonstrate the feasibility of immortalizing short-term cell lines with the SV-40 large T antigen for their use in the characterization of microglial properties.
D000818:Animals; D000831:Animals, Newborn; D000941:Antigens / Q000096:biosynthesis; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis; D001921:Brain / Q000166:cytology*; D002460:Cell Line; D002478:Cells, Cultured; D002999:Clone Cells; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D017628:Microglia / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000941:Antigens; D000952:Antigens, Polyomavirus Transforming; D014409:Tumor Necrosis Factor-alpha; D007770:L-Lactate Dehydrogenase