title: [Effect of the chemically assisted enucleation on the enucleation of sheep oocytes and the development of their reconstructed embryos].
abstract: In order to enhance the efficiency of sheep somatic cell nuclear transfer, we used a chemically assisted enucleation with colchicine to study the effects of the concentration of colchicine, the incubation time of oocytes in colchicine and the maturation time of oocytes on the enucleation rates and the development of reconstructed embryos. The results showed that 1) there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 microg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 microg/mL) for 1 h, and the rate of cytoplast protrusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2) There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 microg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 microg/mL colchicine for 0.5 h. 3) A maturation time of 18-23 h did not affect the rates of cytoplast protrusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4) The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21-23 h. These results demonstrate that sheep oocytes can be enucleated fast and effectively by optimized colcholine chemically assisted enucleation, which can enhance the enucleation rate of sheep oocytes and the early development of reconstructed embryos in vitro.
authors.affiliation: Department of Histology and Embryology, Jilin Medical College, Jilin 132013, China. pxy19790105@163.com
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Pan
authors.forename: Zhengchao
authors.initials: Z
authors.lastname: Wang
authors.forename: Zhixin
authors.initials: Z
authors.lastname: Li
authors.forename: Yuji
authors.initials: Y
authors.lastname: Jin
authors.forename: Zhaohua
authors.initials: Z
authors.lastname: Dou
authors.forename: Zhiqin
authors.initials: Z
authors.lastname: Guo
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000379:methods*; D003078:Colchicine / Q000494:pharmacology*; D004622:Embryo, Mammalian / Q000196:embryology*; D005260:Female; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects; D012756:Sheep
chemical_list: D003078:Colchicine