| Bovine in vitro fertilization(IVF) and embryo transfer (ET) were getting progress greatly during the last two decades in China, the research of embryo biothechniques, such as, bovine somatic cell nuclear transfer(SCNT) , transgenic cattle, developed quickly during the last 10 years. Hence the requirments of bovine oocytes increased, it becomes a very important thing that how to consistently get high quality bovine oocytes for these application and research.Although the oocytes can derived from superovulated cattles,the cost is too expensive. Now main resource of matured bvine oocytes is in vitro matured oocytes derived from ovaries which were collected from slaughter house. In most cases, slaughtering often starts from midnight, and the collection and transport of ovarian often takes several hours, oocytes quality is very easily getting worse during this process and the ovaries collection schedule makes the followed work inconvenient. So in this experiment, addressed the temperature,"biology bao xian ji", and the storage period as factors effecting on the oocytes quality during ovary storage, based on the in vitro maturation rate, parthenogenetic embryos in vitro development potential and the in vivo development potential of SCNT embryos, the bovine ovary storage systems were systemly studied. The experimental results are as follows:1. Low temperature 4℃kept cattle ovarian 0h, 3h, 6h, and 8h, oocyte maturation rate was 69.27%, 67.95% 67.25% and 47.98%, parthenogenetic activation after the cleavage rate was 36.99%, 31.76%, 36.77% and 16.61%, blastocyst rates was 14.16%, 15.56%, 14.43% and 0%, low temperature 4℃preservation 0h,3h and 6h three groups oocyte maturation rate, parthenogenetic activation and the cleavage rate of blastocysts there was no significant differenc(eP>0.05),and low temperature 4℃8h preservation group oocyte maturation rate, cleavage and blastocyst rate was significantly lower than the other three groups(P<0.01);2. Low temperature 4℃add the preservation of biological preservative cattle ovarian 0 h, 3h, 6h, and 8h, oocyte maturation rate of 71.67%, respectively, 69.57%, 68.22% and 67.14% ,Parthenogenetic activation after the cleavage rate was 36.99%, 33.47%, 32.29% and 31.10%, respectively blastocyst 14.63%, 16.43%, 13.89% and 13.53%,Between the four groups was not significant indicators(P>0.05);low temperature 4℃add biological preservative cattle ovarian preservation group and 8h low temperature 4℃kept cattle ovarian 8h group differences between various indicators significantly(P<0.01);3. At room temperature (15 - 20℃) add the preservation of biological preservative cattle ovarian 0h, 3h, 6h, 8h, oocyte maturation rate was 78.53%, 71.32%, 75.16% and 77.98%,parthenogenetic activation after embryo cleavage rate was 38.63%, 36.19%, 33.87% and 33.17%, respectively blastocyst 18.28%, 17.56%, 16.85% and 17.43%,between the four groups was not significant indicators(P>0.05);At room temperature (15 - 20℃) add biology bao xian ji cattle ovarian preservation group and 8h low temperature 4℃add the biology bao xian ji cattle ovarian group 8h between the oocyte maturation rate significantly(0.050.05);4. Maturation of liquid added EGF 0ng/ml, 10ng/ml, 30ng/ml, oocyte maturation rate of 64.86%, respectively, 70.61%, 72.27%,nuclear transfer embryos activated cleavage rate was 60.71%, 80.61%, 82.61%, respectively blastocyst rate 11.88%, 23.81%, 25.00%. maturation of liquid added EGF 10ng/ml group and 30 ng / ml among the indicators there was no significant difference(P>0.05);with the addition of EGF 0ng/ml group differences between various indicators significantly(P<0.01)The results showed that biology bao xian ji can extend the ovary storage period, especially at room temperature (15 - 20℃). 10ng/ml EGF can improve both the in vitro maturation rate and the SCNT embryos development rate.
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