Optimised conditions for the production of hepatitis B virus from cell culture

被引:18
作者
Glebe, D [1 ]
Berting, A [1 ]
Broehl, S [1 ]
Naumann, H [1 ]
Schuster, R [1 ]
Fiedler, N [1 ]
Tolle, TK [1 ]
Nitsche, S [1 ]
Seifer, M [1 ]
Gerlich, WH [1 ]
Schaefer, S [1 ]
机构
[1] Univ Giessen, Inst Med Virol, D-35392 Giessen, Germany
关键词
hepatitis B virus; differentiation; cell culture conditions; hepatocyte;
D O I
10.1159/000050074
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
In chronically infected patients, hepatitis B virus (HBV) particles reach numbers as large as >10(9) genome equivalents (GE)/ml of serum. However, expression of infectious HBV particles in cell culture only yields 10(5)-10(6) GE/ml, which is insufficient for many studies. HBV transcription and possibly replication is dependent on hepatocyte-specific differentiation. Thus, we tested several cell culture parameters that have been reported to enhance the expression of hepatocyte-specific markers, such as growth on different extracellular matrices, different cell culture media, low concentrations of fetal calf serum (FCS) and the addition of dimethyl sulfoxide (DMSO) to the medium. Lower concentrations of FCS, growth on collagen and inclusion of DMSO in the medium only moderately enhanced HBV production in vitro when applied individually. However, combinations of these parameters optimised cell culture conditions and reproducibly increased the release of HBV particles about 100-fold to titres >10(8) GE/ml of culture medium. Copyright (C) 2002 S. Karger AG, Basel.
引用
收藏
页码:370 / 378
页数:9
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