Cloned Guangxi Bama minipig (Sus scrofa) and its offspring have normal reproductive performance

Xenotransplantation is a rapidly expanding field of research, and cloned miniature pigs are considered to be good model animals for its development. Although many animal species have been cloned, the success rate is very low, especially in the pig. To optimize the protocols for somatic cell nuclear transfer in the Guangxi Bama minipig, the relationship between cell cycle synchronization and nuclear histone acetylation levels were investigated. The results showed that the cells were efficiently synchronized by either serum starvation or contact inhibition. The level of nuclear histone acetylation in G0/G1 donor cells had similar variation trends in serum starvation and contact inhibition groups. When the synchronized donor cells were introduced into the enucleated oocytes, 8.8% (serum starvation group) or 9.7% (contact inhibition group) of the reconstructed embryos developed to blastocysts. After embryo transfer, one healthy male Guangxi Bama minipig was obtained. To evaluate the fertility of the cloned pig and its offspring, a series of mating experiments were done. Ninety-eight F1 generation crossbred piglets were born, of which 93 piglets survived. Also, the F1 pigs gave birth to 22 F2 generation piglets, of which 14 piglets survived. In conclusion, a Guangxi Bama minipig was successfully cloned from cultured newborn male gonad fibroblast cells, and the cloned minipig and its offspring had normal fertility.