[Program optimization for bovine somatic cells nuclear transfer].
To optimize program of bovine somatic nuclear transfer, we used two different enucleation procedures (by Spindle-view system & Hoechst 33342 staining), two different procedures to introduce donor nuclei (by ooplasm microinjection & electrofusion), and three different group electrofusion parameters (group 1: 1.9 kV/cm, 10 micros, two; group 2: 1.5 kV/cm, 25 micros, two; group 3: 0.6 kV/cm, 100 micros, one) to reconstruct bovine cloned embryos. The cleavation rates and blastocyst development rates of cloned embryos were used to assess the efficiency of different operational procedure. Finally, the best combination of operational procedure, that the spindle-viewer system was used for oocytes enucleating, and donor cell was electrofused into ooplasm by electrical pulse (1.9 kV/cm, 10 micros, two) to reconstruct bovine cloned embryos. Then the excellent blastocysts were transferred to fosters for producing cloned cattle 80 high-quality cloned blastocysts were transferred into 33 fosters, two cloned calves were produced. According to the results, the optimized program could be used to produce cloned cattle.
Lei
Anmin
A
Shaanxi Stem Cell Engineering and Technology Research Center, Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Veterinary Medicine, Northwest A & F University, Yangling 712100, China. anminleiryan@nwsuaf.edu.cn
Ma
Xiaoling
X
Gao
Zhimin
Z
Hu
Yongce
Y
Sui
Jinqiang
J
Huang
Weiwei
W
Zan
Linsen
L
Dou
Zhongying
Z
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000502:physiology; D019976:Cloning, Organism / Q000662:veterinary*; D004624:Embryo Transfer / Q000379:methods; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D005260:Female; D008845:Microinjections; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology / Q000502:physiology*