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NDT Advance Access published online on July 5, 2006
Nephrology Dialysis Transplantation, doi:10.1093/ndt/gfl327
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© The Author [2006]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received December 2, 2005
Accepted May 9, 2006

Original Article

Targeting of interstitial cells using a simple gene-transfer strategy

Naohiko Fujii 1, Yoshitaka Isaka 2 *, Yoshitsugu Takabatake 1, Masayuki Mizui 1, Chigure Suzuki 1, Shiro Takahara 3, Takahito Ito 1, and Enyu Imai 1

1 Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita 565-0871, Japan
2 Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita 565-0871, Japan; Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan
3 Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine, Suita 565-0871, Japan

* To whom correspondence should be addressed.
Yoshitaka Isaka, E-mail: isaka{at}att.med.osaka-u.ac.jp



  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 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.
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