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
相关论文
共 56 条
[1]   HEPATITIS-B VIRUS PRODUCED BY TRANSFECTED HEP G2 CELLS CAUSES HEPATITIS IN CHIMPANZEES [J].
ACS, G ;
SELLS, MA ;
PURCELL, RH ;
PRICE, P ;
ENGLE, R ;
SHAPIRO, M ;
POPPER, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (13) :4641-4644
[2]   CELL CELL AND CELL MATRIX INTERACTIONS DIFFERENTIALLY REGULATE THE EXPRESSION OF HEPATIC AND CYTOSKELETAL GENES IN PRIMARY CULTURES OF RAT HEPATOCYTES [J].
BENZEEV, A ;
ROBINSON, GS ;
BUCHER, NLR ;
FARMER, SR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (07) :2161-2165
[3]   Effect of extracellular matrix topology on cell structure, function, and physiological responsiveness: Hepatocytes cultured in a sandwich configuration [J].
Berthiaume, F ;
Moghe, PV ;
Toner, M ;
Yarmush, ML .
FASEB JOURNAL, 1996, 10 (13) :1471-1484
[4]  
Bianchi L., 1980, VIRUS LIVER, P197
[5]   THE ROLE OF ENVELOPE PROTEINS IN HEPATITIS-B VIRUS ASSEMBLY [J].
BRUSS, V ;
GANEM, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (03) :1059-1063
[6]   Myristylation of the large surface protein is required for hepatitis B virus in vitro infectivity [J].
Bruss, V ;
Hagelsten, J ;
Gerhardt, E ;
Galle, PR .
VIROLOGY, 1996, 218 (02) :396-399
[7]   Liver-enriched transcription factors and hepatocyte differentiation [J].
Cereghini, S .
FASEB JOURNAL, 1996, 10 (02) :267-282
[8]   PRODUCTION OF HEPATITIS-B VIRUS INVITRO BY TRANSIENT EXPRESSION OF CLONED HBV DNA IN A HEPATOMA-CELL LINE [J].
CHANG, CM ;
JENG, KS ;
HU, CP ;
LO, SCJ ;
SU, TS ;
TING, LP ;
CHOU, CK ;
HAN, SH ;
PFAFF, E ;
SALFELD, J ;
SCHALLER, H .
EMBO JOURNAL, 1987, 6 (03) :675-680
[9]   DIMETHYL-SULFOXIDE ENHANCES LIPID-SYNTHESIS AND SECRETION BY LONG-TERM CULTURES OF ADULT-RAT HEPATOCYTES [J].
DELAVEGA, FM ;
MENDOZAFIGUEROA, T .
BIOCHIMIE, 1991, 73 (05) :621-624
[10]   THE EXTRACELLULAR-MATRIX COORDINATELY MODULATES LIVER TRANSCRIPTION FACTORS AND HEPATOCYTE MORPHOLOGY [J].
DIPERSIO, CM ;
JACKSON, DA ;
ZARET, KS .
MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (09) :4405-4414