Thasler, W. E. and Letschert, K. C. and Weiss, T. and Weinberger, K. M. and Bolder, U. and Jauch, Karl-Walter (1999) Vaccinia mediated gene transfer into human hepatocytes - Efficiency and function under transplant conditions in vitro. LANGENBECKS ARCHIVES OF SURGERY. pp. 247-251. ISSN 1435-2443,
Full text not available from this repository.Abstract
Background/aim: The transplantation of genetically modified livers or hepatocytes has been discussed as therapeutical option for immunomodulation or treatment of liver function defects. A combination of transfection of the gene carrying plasmid and infection with a Vaccinia-Virus in hepatocytes was investigated for a sufficient gene transfer in primary human hepatocytes. The influence of hypothermal storage was evaluated with regard to gene transfer and subsequent gene expression. Methods: Primary human hepatocytes were isolated from human liver resections using EGTA/collagenase-perfusion. After 24 h of culture gene transfer was performed by infection of cells with a recombinant, replication incompetent vaccinia virus followed by liposome-mediated transfection of the reporter plasmid. The gene products were quntified after infection at 37 degrees C and 4 degrees C up to 8 days. Results: Gene transfer was successful in 30% of cells as measured by the expression of gfp (green fluorescent protein) as a marker for transinfection. Under hypothermal conditions gene expression was reduced to 20-25%. The expression of HBsAg in the supernatant was measured up to 8 days of culture. Conclusion: Transinfection with the highly attenuated, replication incompetent vaccinia virus is a valuable model for efficient gene transfer into primary human hepatocytes without major cell damage. Therefore, gene expression of variable plasmids is possible without construction of recombinant vaccinia viral vectors. Organ- or cell preservation at 4 degrees C for potential clinical use in the transplant setting seemes to have no major influence on efficiency and function of gene transfer.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ; gene transfer; vaccinia; human hepatocytes |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Chirurgie Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 30 Nov 2022 09:48 |
| Last Modified: | 30 Nov 2022 09:48 |
| URI: | https://pred.uni-regensburg.de/id/eprint/48686 |
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