Program optimization for bovine somatic cells nuclear transfer.
- Author:
Anmin LEI
1
;
Xiaoling MA
;
Zhimin GAO
;
Yongce HU
;
Jinqiang SUI
;
Weiwei HUANG
;
Linsen ZAN
;
Zhongying DOU
Author Information
1. 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
- Publication Type:Journal Article
- MeSH:
Animals;
Cattle;
Cell Nucleus;
physiology;
Cloning, Organism;
veterinary;
Embryo Transfer;
methods;
Embryo, Mammalian;
cytology;
physiology;
Female;
Microinjections;
Nuclear Transfer Techniques;
veterinary;
Oocytes;
cytology;
physiology
- From:
Chinese Journal of Biotechnology
2009;25(9):1424-1432
- CountryChina
- Language:Chinese
-
Abstract:
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.
