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KMID : 0894520170210020157
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Development & Reproduction
2017 Volume.21 No. 2 p.157 ~ p.165
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Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73
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Lee Seung-Chan
Lee Hae-Sun
Oh Keon-Bong
Hwang In-Sul
Yang Hyeon
Park Mi-Ryung
Ock Sun-A
Woo Jae-Seok
Im Gi-Sun
Hwang Seong-Soo
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Abstract
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One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompati-bility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-human-primate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were con-structed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were 127 ± 18.9. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead af-ter birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachu-setts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as α-1,3-galactosyltransferase knock-out /hCD46 for xenotransplantation.
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KEYWORD
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hCD73 , Porcine Icam2 promoter , Coagulation , Transgenic cloned pig , Xenotransplantation
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