Optimised conditions for the production of hepatitis B virus from cell culture
| Autor: | Maria Seifer, Sylvia Nitsche, Ralph Schuster, Stephan Schaefer, Nicola Fiedler, Dieter Glebe, Wolfram H. Gerlich, Andreas Berting, Tanja K. Tolle, Sigrun Broehl, Heike Naumann |
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| Rok vydání: | 2002 |
| Předmět: |
Hepatitis B virus
Cell Culture Techniques virus diseases Biology medicine.disease_cause Transfection Genome Virology digestive system diseases Microbiology Culture Media Infectious Diseases Equivalent medicine.anatomical_structure Cell culture Hepatocyte medicine Tumor Cells Cultured Humans Dimethyl Sulfoxide Collagen |
| Zdroj: | Intervirology. 44(6) |
| ISSN: | 0300-5526 |
| Popis: | In chronically infected patients, hepatitis B virus (HBV) particles reach numbers as large as >109 genome equivalents (GE)/ml of serum. However, expression of infectious HBV particles in cell culture only yields 105–106 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 >108 GE/ml of culture medium. |
| Databáze: | OpenAIRE |
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