HepG2 Cell Culture Infection by Hepatitis C Virus Genotype 4: Viral Proteins and RNA
HepG2 Cell Culture Infection by Hepatitis C Virus Genotype 4: Viral Proteins and RNA
El-Tabakh, SAA l ; Abdel Wahab, KSE; Badr, AF and Helal, IG
ABSTRACT
Genotype 4 hepatitis C virus (HCV) in viremic serum was used for experimental infection of a human hepatoma cell line (HepG2) at a multiplicity of infection (MOL) of one genome copy per 103 cells. Stock HepG2 cell cultures were grown in Dulbecco modified minimum essential medium (DMMM) as growth medium with 10% fetal calf serum and antibiotic mixture (IOOU penicillin and 100 mgm streptomycin/ml) and subcultured at 3-4 days. HepG2 cells were maintained throughout the experiment in Ham F-12 medium maintenance medium with 2% fetal calf serum; 1000 IU penicillin and 1000 mgm streptomycin antibiotic mixture; with 4% (w/v) polyethylene glycol (PEG), 2% dimethyl sulfoxide (DMSO); and IOYI lovastatin for the experimental infection with HCV. Three markers of HCV virus replication were used. Morphological changes HCV-RNA by reverse transcription polymerase chain reaction (RT PCR) and viral proteins by polyacrylamide gel electrophoresis (PAGE). HCV infected and control HepG2 cells supernatant fluids (SF) were sampled before complete change with fresh MM at weekly intervals for periods extended to one month after infection. Three SF samples taken 5 days apart after HCV infection showed that detection of HCV-RNA in SF was intermittent but' detection of new native protein as well as glycopeptides was consistent.
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July 2008
Vol. 5, Iss. 1
