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Porcine Knock-in Fibroblasts Expressing hDAF on α-1,3-Galactosyltransferase (GGTA1) Gene Locus

  • Kim, Ji-Woo (Department of Animal Science, College of Agriculture and Life Science, Chonnam National University) ;
  • Kim, Hye-Min (Department of Animal Science, College of Agriculture and Life Science, Chonnam National University) ;
  • Lee, Sang-Mi (Department of Animal Science, College of Agriculture and Life Science, Chonnam National University) ;
  • Kang, Man-Jong (Department of Animal Science, College of Agriculture and Life Science, Chonnam National University)
  • Received : 2012.03.17
  • Accepted : 2012.05.01
  • Published : 2012.10.01

Abstract

The Galactose-${\alpha}1$,3-galactose (${\alpha}1$,3Gal) epitope is responsible for hyperacute rejection in pig-to-human xenotransplantation. Human decay-accelerating factor (hDAF) is a cell surface regulatory protein that serves as a complement inhibitor to protect self cells from complement attack. The generation of ${\alpha}1$,3-galactosyltransferase (GGTA1) knock-out pigs expressing DAF is a necessary step for their use as organ donors for humans. In this study, we established GGTA1 knock-out cell lines expressing DAF from pig ear fibroblasts for somatic cell nuclear transfer. hDAF expression was detected in hDAF knock-in heterozygous cells, but not in normal pig cells. Expression of the GGTA1 gene was lower in the knock-in heterozygous cell line compared to the normal pig cell. Knock-in heterozygous cells afforded more effective protection against cytotoxicity with human serum than with GGTA1 knock-out heterozygous and control cells. These cell lines may be used in the production of GGTA1 knock-out and DAF expression pigs for xenotransplantation.

Keywords

Gene Targeting;Pig Fibroblasts;Xenotransplantation;Knock-out;Hyperacute Rejection

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