Serum-free cell culture system supplemented with lipid-rich albumin for hepatitis C virus (strain O of genotype 1b) replication.
HuH-7 is a highly differentiated hepatoma cell line and the only cell line that supports robust RNA replication of the hepatitis C virus (HCV). HuH-7 cells cause cell death in serum-free culture condition. However, the effect is reversed by supplementation with selenium. Serum-free cell cultures are advantageous for vaccine development and experimental reproducibility. However, HCV RNA replication in HuH-7 cells in serum-free medium had not yet been achieved. Therefore, we tried to develop a system for robust HCV RNA replication in a serum-free cell culture. Although HuH-7 cells grew in serum-free medium in the presence of selenium, HuH-7 cells under these conditions did not support HCV RNA replication in long-term culture. Among the supplements tested, serum-free medium with lipid-rich albumin (LRA) was found to yield robust HCV RNA replication. HCV proteins were detected for more than 9 months in serum-free medium supplemented with LRA. This is the first report to demonstrate a long-term, serum-free cell culture that successfully maintained robust HCV RNA replication. This cell culture system is expected to be a useful tool for vaccine development, as well as for further investigation of cellular factors that are essential for HCV RNA replication.
Abe
Ken-ichi
K
Department of Molecular Biology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1, Shikata-cho, Okayama 700-8558, Japan.
Ikeda
Masanori
M
Ariumi
Yasuo
Y
Dansako
Hiromichi
H
Kato
Nobuyuki
N
D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005312:Fetal Blood; D016174:Hepacivirus / Q000145:classification / Q000502:physiology*; D006801:Humans; D012367:RNA, Viral / Q000096:biosynthesis; D012643:Selenium / Q000494:pharmacology; D012709:Serum Albumin / Q000494:pharmacology*; D014779:Virus Replication
D000998:Antiviral Agents; D016895:Culture Media, Serum-Free; D012367:RNA, Viral; D012709:Serum Albumin; D012643:Selenium