RT Journal Article SR Electronic(1) A1 Seipp, Stefanie A1 Mueller, Hubert M A1 Pfaff, Eberhard A1 Stremmel, Wolfgang A1 Theilmann, Lorenz A1 Goeser, TobiasYR 1997 T1 Establishment of persistent hepatitis C virus infection and replication in vitro. JF Journal of General Virology, VO 78 IS 10 SP 2467 OP 2476 DO https://doi.org/10.1099/0022-1317-78-10-2467 PB Microbiology Society, SN 1465-2099, AB Hepatitis C virus (HCV) is a major cause of chronic viral hepatitis. Development of anti-viral strategies has been hampered by the lack of efficient cell systems to propagate HCV in vitro. To establish a long-term culture system, we tested human hepatoma (HuH7, HepG2) and porcine non-hepatoma (PK15, STE) cell lines, as well as several culture and infection conditions. As a marker for virus replication, minus-strand HCV RNAin infected cells was detected by an enhanced detection system using nested RT-PCR followed by hybridization analysis. Short-term efficiency of HCV infection (10 days) was slightly increased by addition of polyethylene glycol (PEG) and/or dimethyl sulfoxide (DMSO) to culture media during inoculation of HuH7, PK15 and STE cells, but no augmentation in long-term culture was achieved, suggesting enhanced attachment of HCV to cells rather than more efficient infection. A stabilizing effect on HCV propagation was observed for 50 days in a serum-free medium with stimulation of the low-density lipoprotein (LDL) receptor expression by lovastatin. Using partially serum-free culture conditions, long-term persistence of HCV in cells and release of virions into supernatant was achieved for up to 130 days. Infectivity of released virions in supernatants after long-term culturing (day 30–80) was shown by successful infection of fresh cells. In conclusion, supplementation with PEG, DMSO and lovastatin during inoculation did not enhance virus replication substantially, but continued stimulation of LDL-receptor expression resulted in infections which persisted for over 4 months. These data support the hypothesis of an LDL-receptor mediated uptake of HCV into cells in vitro., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-78-10-2467