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Establishment of normal and lethal albino deletion mutant mouse hepatocyte lines immortalized by SV40 virus

: Kwon, B.S.; Hoffmann, B.; Höhne, M.; Kay, G.; Paul, D.; Tönjes, R.

Cold Spring Harbor Laboratory, New York/NY:
Meeting on Regulation of Liver Gene Expression 1989. Abstracts of papers
New York/N.Y., 1989
Meeting on Regulation of Liver Gene Expression <1989, Cold Spring Harbor/N.Y.>
Conference Paper
Fraunhofer ITA ( ITEM) ()
hepatocyte; liver; mouse; SV 40 virus

Primary liver cultures of newborn albino deletion mutant mice lacing about 1.5 Kb on chromosome 7 at the albino locus (c) (c high 14Cos/c high 14Cos) and of phenotypically normal mice (c high 14Cos/c high ch or c high ch) (Gluecksohn-Waelsch, Cell 16: 225-237 (1979) were infected with SV40-virus. Resulting multiplying hepatocytes were subcultured by selection in arginine-deficient serum-free medium containing EFG, insulin and hydrocortisone (HC) (Hoffmann et al., J.Cell.Physiol. (in press)). Resulting normal (NMH-ch) and mutant (NMH-m14) hepatocyte lines expressing intregrated viral transforming sequences did not senesce, multiplied in medium with insulin plus HC, have abrogated EGF requirements, and retained hepatocyte-specific functions. Both lines synthesized arginine and contained albumin and alpha-fetoprotein (AFP) mRNA's. HC-inducible tyrosine-amino-tranferase (TAT)-specific mRNA was detected in normal but not in mutant hepatocyte lines. Mutant hepatocyte lines were characterized by Southern analysis using cDNA sequences encoding human tyrosinase, known to map at the mouse albino locus (Kwon et al., PNAS 84: 7473-7477 (1987) within sequences corresponding to the deleted region in the c high 14Cos mutant. Tyrosinase cDNA sequences, hybridized with a 2.5 kb EcoRI fragment of normal NMH-ch DNY, whereas this fragment did not hybridize with mutant NMH-m14 DNA.