Characterization and immortalization of woodchuck hepatocytes isolated from normal and hepadnavirus-infected woodchucks (Marmota monax).
Primary woodchuck (Marmota monax) hepatocytes from normal woodchucks and woodchucks with chronic woodchuck hepatitis virus (WHV) infection were cultured in either a conventional serum-containing medium or a serum-free medium. The de novo synthesis of the plasma proteins albumin, transferrin, fibrinogen, and complement C3 were identical under both conditions. However, expression of the WHV and the synthesis of nitric oxide were diminished under serum-free conditions. Primary woodchuck hepatocytes cultured in conventional, serum-containing medium were immortalized utilizing the simian virus 40 T antigen oncogene. Immortalized hepatic cell lines retained differentiated functions of nitric oxide synthesis and expression of complement C3. The woodchuck hepatocyte culture model will supplement current experimental methods, allowing investigation of hepadnaviral pathogenesis, including hepatocarcinogenesis in vitro.
Jacob
J R
JR
College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
Liu
R H
RH
Roneker
C A
CA
de Noronha
F
F
Hotchkiss
J H
JH
Tennant
B C
BC
D000418:Albumins / Q000032:analysis; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000209:etiology / Q000382:microbiology; D002455:Cell Division; D002461:Cell Line, Transformed / Q000382:microbiology*; D002469:Cell Separation; D002472:Cell Transformation, Viral; D003176:Complement C3 / Q000032:analysis; D016895:Culture Media, Serum-Free; D005340:Fibrinogen / Q000032:analysis; D006509:Hepatitis B / Q000382:microbiology*; D017919:Hepatitis B Virus, Woodchuck / Q000254:growth & development*; D008099:Liver / Q000166:cytology* / Q000382:microbiology; D008392:Marmota; D009569:Nitric Oxide / Q000096:biosynthesis; D013539:Simian virus 40 / Q000235:genetics; D014168:Transferrin / Q000032:analysis
D000418:Albumins; D000957:Antigens, Viral, Tumor; D003176:Complement C3; D016895:Culture Media, Serum-Free; D014168:Transferrin; D009569:Nitric Oxide; D005340:Fibrinogen