JCI - Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection

Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection

Xiping Zhao, Zhen-Ya Tang, Bettina Klumpp, Guido Wolff-Vorbeck, Heidi Barth, Shoshana Levy, Fritz von Weizsäcker, Hubert E. Blum, Thomas F. Baumert

Published in Volume 109, Issue 2

J Clin Invest. 2002; 109(2):221–232 doi:10.1172/JCI13011

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Figure 2

Passage of HCV in primary Tupaia hepatocytes. (a) Passage of HCV to naive cells. Primary Tupaia hepatocytes were incubated with HCV RNA–positive plasma or serum (1: 10 μl H77 7-12-77, 10^–1 dilution; 2: 10 μl ES-2; or 3: 50 μl A5387) 1 day after plating. 5 days later, HCV infection was determined by RT-PCR of cellular positive- and negative-strand HCV RNA, on day 5 after incubation with serum-derived HCV, as described above. Strand-specificity of RT-PCR was assessed using 10 fg of in vitro–synthesized positive- (lanes 1 and 7) and negative-strand HCV RNA (lanes 2 and 8). On day 5 after infection, medium was transferred to naive hepatocytes, and infection was assessed 5 days later as described above. (b) Serial passage. Primary Tupaia hepatocytes were infected with serum 3 as described above. On day 5 after infection, medium was transferred to naive hepatocytes from a separate preparation (first passage). Eight days later, medium was used to infect a third preparation of hepatocytes (second passage). Infection of hepatocytes was assessed as described above.