Parthenogenetic Activation of Bovine Oocytes and Heterogeneous Embryo Reconstruction by Horse Skin Fibroblasts and Bovine Oocytes

Abstract: Firstly， in vitro matured bovine oocytes were activated by three different activators. The results showed that matured bovine oocytes were efficiently activated by Inomycin， A23187 and 7% ethanol alone followed by 6-DMAP， respectively， and developed to blastocyst phase， but the cleavage rate and the blastocyst rate of matured bovine oocytes activated by inomycin-6-DMAP were significantly higher than those of matured bovine oocytes activated by the two others (P<0.05); When the activated oocytes were cultured in SOFaa added with 10% FBS and cumuluses， a higher cleavage rate and blastocyst rate were achieved (72.30%， 14.91%). Secondly， horse skin fibroblasts were cultured in vitro and induced to quiescence stage (G0 phase) by serum starvation culture. The cells were injected into enucleated bovine oocytes and the reconstructed heterogeneous embryos could be obtained. The cell fusion was induced by 2.0 kV/cm for 40 µs under the condition of 20 V start-voltage， 10 s duration， 0.2 MHz frequency， 15 V end-voltage and an interval of 0.125 s， fusion rate and cleavage rate of reconstructed embryos reached the peak values (52.27%， 71.74%).

CSTR: 32200.14.cjcb.2006.06.0023