Cultivation of Hepatitis Virus in Tissue Culture.
Annual rept. no. 5, 1 Mar 78-28 Feb 79,
CALIFORNIA UNIV LOS ANGELES DEPT OF MEDICINE
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Since hepatitis virus has the propensity in vivo to infect human hepatocytes it seemed appropriate to undertake attempts at hepatitis virus cultivation using hepatocyte cultures. Early studies led to technology for the cultivation of fetal hepatocytes with the exclusion of fibroblastic cells. Similar approaches were used to cultivate adult hepatocytes and most recently these recent studies have demonstrated the successful long-term cultivation of adult hepatocytes for periods in excess of 10 weeks. The conditions for growth have been delineated and the methods are now reproducible. In the past twelve months, attempts were made to use hepatocyte cultures as a substrate for hepatitis B and hepatitis A cultivation. The inocula used have been hepatitis B antigen positive sera known to be rich in Dane particles, filtrates of stools from patients with hepatitis A and liver biopsies obtained from patients with hepatitis B surface antigen positive chronic aggressive hepatitis. Inoculation of cultures with Dane particle-rich hepatitis B sera has not resulted in cytopathogenic change CPE in monolayer cultures. Efforts have been devoted at attempts at cultivation of hepatitis B virus by using slow virus techniques. By means of cocultivation, mixed monolayers of normal hepatocytes were co-cultivated with hepatocytes obtained from liver biopsies of patients with hepatitis B surface antigen positive chronic aggressive hepatitis. The cell layers which resulted were maintained and were sub-cultured but did not yield evidence of either CPE or evidence of multiplication of hepatitis B surface antigen.