An oligodendrocyte precursor cell line from rat optic nerve.
We have established permanent cell lines from the optic nerve of the rat with a temperature sensitive immortalizing oncogene (Simian Virus 40 large T-antigen carrying both the tsA58 and U19 mutations). The oncogene was transduced into primary cultures via a replication deficient retrovirus, and infected cells were selected with the antibiotic G418. A clonal cell line (tsU19-5) displayed some properties of oligodendrocyte precursors: it proliferated, bound the monoclonal antibody A2B5 (which recognizes minor ganglioside species), and expressed the intermediate filament vimentin and the enzyme 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNP) at 33 degrees C (the permissive temperature for the oncogene). At 39 degrees C (the non-permissive temperature), some cells had the potential to differentiate further, and expressed several oligodendrocyte specific components: galactocerebroside, myelin basic protein, proteolipid protein and CNP. These results suggest that conditional oncogenes can establish neural precursor cell lines which are still capable of differentiation in vitro.
Almazan
G
G
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge 02139.
McKay
R
R
D000818:Animals; D015139:Blotting, Southern; D001973:Bromodeoxyuridine / Q000378:metabolism; D002460:Cell Line; D002999:Clone Cells; D005455:Fluorescent Antibody Technique; D008854:Microscopy, Electron; D009836:Oligodendroglia / Q000502:physiology* / Q000648:ultrastructure; D009900:Optic Nerve / Q000166:cytology*; D010957:Plasmids; D012367:RNA, Viral / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D012190:Retroviridae / Q000235:genetics; D013936:Thymidine / Q000378:metabolism; D014162:Transfection
D012367:RNA, Viral; D001973:Bromodeoxyuridine; D013936:Thymidine