High efficiency gene transfer into cultured primary rat and human hepatic stellate cells using baculovirus vectors
- PMID: 11906614
- DOI: 10.1046/j.0106-9543.2001.01555.x
High efficiency gene transfer into cultured primary rat and human hepatic stellate cells using baculovirus vectors
Abstract
Background/aims: Gene transfer into hepatic stellate cells (HSC) is inefficient when using plasmid-based transfection methods; viral-based systems are therefore being developed. A baculovirus system has recently been shown to be useful for expressing genes in mammalian cells. The aim of this study was to determine if baculovirus vectors can infect and express target genes in rat and human HSC and to assess potential cytotoxic and modulatory effects of infection.
Methods: A recombinant baculovirus vector (AcCALacZ) carrying the LacZ gene was used to infect HSC. beta-Galactosidase assays and electron microscopy were used to determine efficiency of infection and gene expression. Counting of trypan blue negative cells was used to assess cytotoxic/cytostatic effects of infection. Measurement of protein content of cells and alpha-smooth muscle actin expression were performed to assess the effects of baculovirus on cell function/phenotype.
Results: Baculovirus infection of activated HSC was highly efficient (> 90%) and provided long-term LacZ gene expression (15 days) in the absence of cytotoxic, cytostatic or modulatory effects. Infection of freshly isolated cells was also observed but at lower levels (20%).
Conclusions: Baculovirus vectors can therefore be used to deliver target genes to cultured rat and human HSC with high efficiency and longevity in the absence of detrimental effects on cell function.
Similar articles
-
Efficient transduction of mammalian cells by a recombinant baculovirus having the vesicular stomatitis virus G glycoprotein.Hum Gene Ther. 1997 Nov 20;8(17):2011-8. doi: 10.1089/hum.1997.8.17-2011. Hum Gene Ther. 1997. PMID: 9414250
-
Baculovirus vector requires electrostatic interactions including heparan sulfate for efficient gene transfer in mammalian cells.J Gene Med. 1999 Mar-Apr;1(2):93-102. doi: 10.1002/(SICI)1521-2254(199903/04)1:2<93::AID-JGM19>3.0.CO;2-1. J Gene Med. 1999. PMID: 10738573
-
Potential usefulness of baculovirus-mediated sodium-iodide symporter reporter gene as non-invasively gene therapy monitoring in liver cancer cells: an in vitro evaluation.Technol Cancer Res Treat. 2014 Apr;13(2):139-48. doi: 10.7785/tcrt.2012.500368. Epub 2013 Aug 2. Technol Cancer Res Treat. 2014. PMID: 23919394
-
Baculovirus vectors: novel mammalian cell gene-delivery vehicles and their applications.Am J Pharmacogenomics. 2003;3(1):53-63. Am J Pharmacogenomics. 2003. PMID: 12562216 Review.
-
Towards the use of baculovirus as a gene therapy vector.Curr Opin Mol Ther. 2001 Oct;3(5):464-7. Curr Opin Mol Ther. 2001. PMID: 11699890 Review.
Cited by
-
Baculovirus as a gene delivery vector: recent understandings of molecular alterations in transduced cells and latest applications.Biotechnol Adv. 2011 Nov-Dec;29(6):618-31. doi: 10.1016/j.biotechadv.2011.04.004. Epub 2011 Apr 28. Biotechnol Adv. 2011. PMID: 21550393 Free PMC article. Review.
-
Baculovirus vectors for gene therapy.Adv Virus Res. 2006;68:287-320. doi: 10.1016/S0065-3527(06)68008-1. Adv Virus Res. 2006. PMID: 16997015 Free PMC article. Review.
-
Baculovirus as a highly efficient expression vector in insect and mammalian cells.Acta Pharmacol Sin. 2005 Apr;26(4):405-16. doi: 10.1111/j.1745-7254.2005.00078.x. Acta Pharmacol Sin. 2005. PMID: 15780188 Free PMC article. Review.
-
Enhanced baculovirus-mediated transduction of human cancer cells by tumor-homing peptides.J Virol. 2006 Jul;80(13):6603-11. doi: 10.1128/JVI.00528-06. J Virol. 2006. PMID: 16775347 Free PMC article.
-
Baculovirus as versatile vectors for protein expression in insect and mammalian cells.Nat Biotechnol. 2005 May;23(5):567-75. doi: 10.1038/nbt1095. Nat Biotechnol. 2005. PMID: 15877075 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
