The ability of hepatitis C virus (HCV) to replicate in two B-cell lines, CE and TOFE, derived from bone marrow of healthy subjects was compared using qualitative and quantitative molecular methods. The presence of intracellular negative-stranded HCV RNA (replicative intermediate) was investigated by nested polymerase chain reaction (PCR) in the infected cultures at different times after infection. The amounts of positivestranded HCV RNA (genomic RNA copies) synthesized and released from cells one week after in vitro infection were determined by competitive PCR after reverse transcription of viral RNA for the 5′ viral untranslated region. In both cell lines, HCV RNA replication took place, but the TOFE cell line appeared to be a more efficient virus producer than the CE cell line. The TOFE cell line could be a valuable and reliable tool for basic and clinical HCV studies.