title: Targeting of interstitial cells using a simple gene-transfer strategy.
abstract: BACKGROUND
  Interstitial fibroblasts are central to the inflammatory response during the progression of tubulointerstitial fibrosis. We examined the efficiency of a new gene transfer method that targets interstitial cells by using parenchymal injection of DNA followed by electroporation.
  
  
  METHODS
  Fluoresceinisothiocyanate-labelled oligodeoxynucleotides (FITC-ODNs) or expression vectors were directly injected into the cortex of the kidney, followed by electroporation.
  
  
  RESULTS
  Transfection with FITC-ODNs or the EGFP expression vector resulted in efficient transfection in interstitial fibroblasts, but not in tubular epithelial cells or glomerular cells. Transfection efficiency was optimal after using a total of 150 microg of DNA in 1000 microl of PBS, combined with clamping of the renal vessels prior to electroporation. Gene expression peaked at 4 days after transfection and decreased by two orders of magnitude at 6 weeks post-transfection; however, expression recovered to near peak levels after parenchymal or intraperitoneal injection of FR901228, a histone deacetylase inhibitor.
  
  
  CONCLUSIONS
  We demonstrated that direct parenchymal injection of DNA combined with electroporation enables gene transfer into interstitial fibroblasts.
authors.affiliation: Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.
authors.forename: Naohiko
authors.initials: N
authors.lastname: Fujii
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Isaka
authors.forename: Yoshitsugu
authors.initials: Y
authors.lastname: Takabatake
authors.forename: Masayuki
authors.initials: M
authors.lastname: Mizui
authors.forename: Chigure
authors.initials: C
authors.lastname: Suzuki
authors.forename: Shiro
authors.initials: S
authors.lastname: Takahara
authors.forename: Takahito
authors.initials: T
authors.lastname: Ito
authors.forename: Enyu
authors.initials: E
authors.lastname: Imai
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology; D003239:Connective Tissue Cells / Q000187:drug effects / Q000378:metabolism; D047630:Depsipeptides / Q000008:administration & dosage / Q000494:pharmacology; D018274:Electroporation / Q000379:methods; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage / Q000737:chemistry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014162:Transfection / Q000379:methods
chemical_list: D000903:Antibiotics, Antineoplastic; D047630:Depsipeptides; D009838:Oligodeoxyribonucleotides; D049452:Green Fluorescent Proteins; C087123:romidepsin; D008156:Luciferases; D016650:Fluorescein-5-isothiocyanate