title: High efficiency gene transfer into cultured primary rat and human hepatic stellate cells using baculovirus vectors.
abstract: OBJECTIVE
  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.
authors.affiliation: Liver Group, Division of Infection, Inflammation and Repair, University of Southampton, Level D, South Academic Block, Southampton General Hospital, Tremona Road, Southampton, UK.
authors.forename: Runping
authors.initials: R
authors.lastname: Gao
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: McCormick
authors.forename: Michael J P
authors.initials: MJ
authors.lastname: Arthur
authors.forename: Richard
authors.initials: R
authors.lastname: Ruddell
authors.forename: Fiona
authors.initials: F
authors.lastname: Oakley
authors.forename: David E
authors.initials: DE
authors.lastname: Smart
authors.forename: Frank R
authors.initials: FR
authors.lastname: Murphy
authors.forename: Mark P G
authors.initials: MP
authors.lastname: Harris
authors.forename: Derek A
authors.initials: DA
authors.lastname: Mann
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000648:ultrastructure; D002455:Cell Division / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007763:Lac Operon; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D011995:Recombination, Genetic; D018411:Spodoptera
chemical_list: D000199:Actins