Production of human CD59-transgenic pigs by embryonic germ cell nuclear transfer.
This study was performed to produce transgenic pigs expressing the human complement regulatory protein CD59 (hCD59) using the nuclear transfer (NT) of embryonic germ (EG) cells, which are undifferentiated stem cells derived from primordial germ cells. Because EG cells can be cultured indefinitely in an undifferentiated state, they may provide an inexhaustible source of nuclear donor cells for NT to produce transgenic pigs. A total of 1980 NT embryos derived from hCD59-transgenic EG cells were transferred to ten recipients, resulting in the birth of fifteen piglets from three pregnancies. Among these offspring, ten were alive without overt health problems. Based on PCR analysis, all fifteen piglets were confirmed as hCD59 transgenic. The expression of the hCD59 transgene in the ten living piglets was verified by RT-PCR. Western analysis showed the expression of the hCD59 protein in four of the ten RT-PCR-positive piglets. These results demonstrate that hCD59-transgenic pigs could effectively be produced by EG cell NT and that such transgenic pigs may be used as organ donors in pig-to-human xenotransplantation.
Ahn
Kwang Sung
KS
Department of Physiology, Dankook University School of Medicine, Cheonan, Republic of Korea.
Won
Ji Young
JY
Park
Jin-Ki
JK
Sorrell
Alice M
AM
Heo
Soon Young
SY
Kang
Jee Hyun
JH
Woo
Jae-Seok
JS
Choi
Bong-Hwan
BH
Chang
Won-Kyong
WK
Shim
Hosup
H
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D018957:CD59 Antigens / Q000235:genetics*; D004622:Embryo, Mammalian / Q000166:cytology*; D005854:Germ Cells / Q000378:metabolism*; D006801:Humans; D053652:Nuclear Transfer Techniques; D013552:Swine / Q000235:genetics*
D018957:CD59 Antigens; C051590:CD59 protein, human