Developmental competence of somatic cell nuclear transfer embryos reconstructed from oocytes matured in vitro with follicle shells in miniature pig.
The developmental competence of domestic pig oocytes that were transferred to somatic cell nuclei of miniature pig was examined. A co-culture system of oocytes with follicle shells was used for the maturation of domestic pig oocytes in vitro. Co-cultured oocytes progressed to the metaphase II stage of meiosis more quickly and more synchronously than non co-cultured oocytes. Oocytes were enucleated and fused with fibroblast cells of Potbelly miniature pig at 48 h of maturation. The blastocyst formation rate of nuclear transfer (NT) embryos using cocultured oocytes (24%) was significantly higher (p < 0.05) than that of non-co-cultured oocytes (13%). Cleaved embryos at 48 h after nuclear transfer using co-cultured oocytes were transferred to the oviducts of 14 Göttingen miniature pigs and four Meishan pigs. Estrus of all Göttingens returned at around 20-31 days of pregnancy. Two of the four Meishans became pregnant. Three and two cloned piglets were born after modest number of embryo transfer (15 and 29 embryos transferred), respectively. These results indicated that oocytes co-cultured with follicle shells have a high developmental competence after nuclear transfer and result in full-term development after embryo transfer.
Hoshino
Yoichiro
Y
Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan.
Uchida
Masaki
M
Shimatsu
Yoshiki
Y
Miyake
Masashi
M
Nagao
Yasumitsu
Y
Minami
Naojiro
N
Yamada
Masayasu
M
Imai
Hiroshi
H
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism / Q000379:methods*; D018920:Coculture Techniques; D046508:Culture Techniques; D000242:Cyclic AMP / Q000378:metabolism; D004624:Embryo Transfer; D047108:Embryonic Development; D004971:Estrus; D005260:Female; D007668:Kidney / Q000166:cytology; D008168:Lung / Q000166:cytology; D008297:Male; D008540:Meiosis; D008677:Metaphase; D018895:Microsatellite Repeats; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D013552:Swine; D013556:Swine, Miniature; D013997:Time Factors
D000242:Cyclic AMP