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Develeopment of Bovine Somatic Cell Nuclear Transfer and
Parthenogenesis Embryos in Two-step Culture System In vitro |
| LI Yu-Qiang ZHANG Xiu LI Yu ZHANG Yong |
| (Animal Sci-tech College, Northwest Sci-tech University of Agriculture and Forestry, Yangling 712100, China) |
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Abstract Abstract: The development of bovine parthenogenesis and somatic cell nuclear transfer embryos in different culture systems was studied. In vitro maturated bovine oocytes were activated by 5 mmol/L ionomycin for 5 min treatment and 2 mmol/L 6-DMAP for 4 h culture. The nuclear transplantation embryos were produced by intracytoplasmic injection of the bovine skin fibroblast nuclear. The results indicated that the blastocyte rate was higher when bovine parthenogenesis embryos cultured in BECMaa (bovine embryo culture media) + 10% FBS than in SOFaa + 10% FBS or TCM199 + 10% FBS (16.5%∶13.6% / 12.8%, P＜0.05); However, adding 1 mmol/L glutathione into SOFaa + 10% FBS could significantly increase parthenogenesis blastocyte rate (18.5%∶12.8%, P＜0.01). When added into serum-free SOFaa (including GSH) the fibroblast growth factor 4 (FGF4) and insulin could improve bovine nuclear transfer embryo development significantly respectively (8.7% / 9.3%:5.5%, P ＜0.05), but there was no obviously effect when FGF4 was used together with FBS. According to the above results and metabolism character of early bovine embryo, the two-step bovine embryo culture system in vitro, which embryos were cultured firstly in media Ⅰ(SOFnaa + EDTA + GSH + insulin) for 72 h, then transferred into media Ⅱ (SOFaa + 5％FBS + Glucose + GSH + insulin) for 8 d, was established. As compared with one-step culture using media SOFaa + 10％FBS + GSH, two-step culture system could significantly improve bovine parthenogenesis and nuclear transfer embryos development, whichever the blastocyte rate (22.3%∶18.5%; 15.0%∶11.7%, P＜0.01) or the development rate from 8/16-cell stage to blastocyte(47.3%∶43.6%; 37.4%∶32.5%, P＜0.05). In conclusion, the two-step culture system was a feasible bovine embryos culture method in vitro, and much more accorded with the development character and nutrition demand of bovine early embryo.
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Received: 01 January 1900
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Corresponding Authors:
LI Yu-Qiang
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