NDT Advance Access originally published online on July 5, 2006
Nephrology Dialysis Transplantation 2006 21(10):2745-2753; doi:10.1093/ndt/gfl327
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Targeting of interstitial cells using a simple gene-transfer strategy
1Department of Internal Medicine and Therapeutics and 2Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan
Correspondence and offprint requests to: Yoshitaka Isaka, MD, PhD, Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan. Email: isaka{at}att.med.osaka-u.ac.jp
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 µg of DNA in 1000 µl 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.
Conclusion. We demonstrated that direct parenchymal injection of DNA combined with electroporation enables gene transfer into interstitial fibroblasts.
Keywords: electroporation; gene transfer; interstitial fibroblast; parenchymal