Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Characterization of α-Gal Epitope in Cells and Tissues from Homozygous α-1,3-Galactosyltransferase Knockout Pigs

  • Hwang, In-Sul (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kwon, Dae-Jin (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kwak, Tae-Uk (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Oh, Keon Bong (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Ock, Sun-A (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Hak-Jae (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Hwang, Seongsoo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2015.11.10
  • Accepted : 2015.11.18
  • Published : 2015.12.31
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Abstract

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred ${\alpha}$-1,3-galactosyltransferase knock-out ($GalT^{-/-}$) pigs. In this study, the somatic cells and tissues from the $GalT^{-/-}$ pigs were characterized by an analysis of the expression of Gal${\alpha}$-1,3-Gal (${\alpha}-Gal$) epitope. Briefly, ear fibroblast cell lines of 19 homozygous $GalT^{-/-}$ pigs were established and cryopreserved. The expression of ${\alpha}-Gal$ epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous ($GalT^{-/-}$) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of ${\alpha}-Gal$ epitope expression. The results showed that the expression of ${\alpha}-Gal$ epitope in $GalT^{-/-}$ cells (0.2 %) were significantly (p<0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous ($GalT^{-/+}$) (9.3 %) and wild type ($GalT^{+/+}$) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of ${\alpha}-Gal$ epitope was detected a partly in $GalT^{-/+}$ cells and mostly in $GalT^{+/+}$ cells, it was almost not detected in the $GalT^{-/-}$ cells. Also, immunostaining results from various tissues of the $GalT^{-/-}$ pig showed that the expression of ${\alpha}-Gal$ epitope was not detectable, whereas various tissues from $GalT^{+/+}$ pig showed a strong expression of ${\alpha}-Gal$ epitope. Our results demonstrated that ${\alpha}-Gal$ epitope expressions from $GalT^{-/-}$ pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

Keywords

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