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Immortalization and characterization of rat microglial cells
Author(s) -
McLaurin J.,
Almazan G.,
Williams K.,
Antel J. P.
Publication year - 1995
Publication title -
neuropathology and applied neurobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.538
H-Index - 95
eISSN - 1365-2990
pISSN - 0305-1846
DOI - 10.1111/j.1365-2990.1995.tb01064.x
Subject(s) - antigen , biology , cell culture , microglia , microbiology and biotechnology , major histocompatibility complex , phagocytosis , mhc class ii , macrophage , granulocyte , immunology , virology , in vitro , biochemistry , genetics , inflammation
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 proIiferate actively at 33°C. whereas the rate of division was significantly decreased at 39°C when the large T antigen is non‐functional. At 39°C these cells take on the microglial phenotype as demonstrated by immunoreactivity to ED‐1 (an intra‐cellular 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 (γ‐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.