To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred α-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 Galα-1,3-Gal (α-Gal) epitope. Briefly, ear fibroblast cell lines of 19 homozygous GalT —/— pigs were established and cryopreserved. The expression of α-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 α-Gal epitope expression. The results showed that the expression of α-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 α-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 α-Gal epitope was not detectable, whereas various tissues from GalT +/+ pig showed a strong expression of α-Gal epitope. Our results demonstrated that α-Gal epitope expressions from GalT —/— pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

• In-Sul Hwang(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Dae-Jin Kwon(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Tae-Uk Kwak(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Keon Bong Oh(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Sun-A Ock(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Hak-Jae Chung(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Gi-Sun Im(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,)
• Seongsoo Hwang(Animal Biotechnology Division, National Institute of Animal Science,Rural Development Administration,) †Corresponding author