Cloning of Asian yellow goat (C. hircus) by somatic cell nuclear transfer: telophase enucleation combined with whole cell intracytoplasmic injection.
Our and other previous studies have shown that telophase enucleation is an efficient method for preparing recipient cytoplasts in nuclear transfer. Conventional methods of somatic cell nuclear transfer either by electro-fusion or direct nucleus injection have very low efficiency in animal somatic cell cloning. To simplify the manipulation procedure and increase the efficiency of somatic cell nuclear transfer, this study was designed to study in vitro and in vivo development of Asian yellow goat cloned embryos reconstructed by direct whole cell intracytoplasmic injection (WCICI) into in vitro matured oocytes enucleated at telophase II stage. Our results demonstrated that the rates of cleavage and blastocyst development of embryos reconstructed by WCICI were slightly higher than in conventional subzonal injection (SUZI) group, but no statistic difference (P > 0.05) existed between these two methods. However, the percentage of successful embryonic reconstruction in WCICI group was significantly higher than that in SUZI group (P < 0.05). After embryo transfer at 4-cell stage, the foster in both groups gave birth to offspring. Therefore, the present study suggests that the telophase ooplasm could properly reprogram the genome of somatic cells, produce Asian yellow goat cloned embryos and viable kids, and whole cell intracytoplasmic injection is an efficient protocol for goat somatic cell nuclear transfer.
Chen
Da-Yuan
DY
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Haidian, Beijing, China. chendy@ioz.ac.cn
Jiang
Man-Xi
MX
Zhao
Zhen-Jun
ZJ
Wang
Hai-Long
HL
Sun
Qing-Yuan
QY
Zhang
Li-Sheng
LS
Li
Rui-Chang
RC
Cao
Heng-Hua
HH
Zhang
Quan-Jun
QJ
Ma
Dong-Lian
DL
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism / Q000379:methods*; D003593:Cytoplasm / Q000378:metabolism; D006041:Goats / Q000235:genetics*; D008845:Microinjections; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000378:metabolism; D013692:Telophase