title: Optimised conditions for the production of hepatitis B virus from cell culture.
abstract: 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.
authors.affiliation: Institut für Medizinische Virologie, Justus-Liebig-Universität, Giessen, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Glebe
authors.forename: A
authors.initials: A
authors.lastname: Berting
authors.forename: S
authors.initials: S
authors.lastname: Broehl
authors.forename: H
authors.initials: H
authors.lastname: Naumann
authors.forename: R
authors.initials: R
authors.lastname: Schuster
authors.forename: N
authors.initials: N
authors.lastname: Fiedler
authors.forename: T K
authors.initials: TK
authors.lastname: Tolle
authors.forename: S
authors.initials: S
authors.lastname: Nitsche
authors.forename: M
authors.initials: M
authors.lastname: Seifer
authors.forename: W H
authors.initials: WH
authors.lastname: Gerlich
authors.forename: S
authors.initials: S
authors.lastname: Schaefer
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods / Q000592:standards*; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006515:Hepatitis B virus / Q000187:drug effects / Q000254:growth & development*; D006801:Humans; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D003470:Culture Media; D003094:Collagen; D004121:Dimethyl Sulfoxide