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Methodlogical Studies of Cryopreservation of Bovine Oocytes and Embryos

Author: WangZuo
Tutor: ZhaoXingXu
School: Gansu Agricultural University
Course: Clinical Veterinary Medicine
Keywords: Bovine oocytes embryos cryopreservation
CLC: S823
Type: Master's thesis
Year: 2008
Downloads: 48
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Abstract


Cryopreservation of oocytes and embryos is a potentially valuable technique for conservation of farm animal genetic resource and population. The bovine oocytes were vitrified by different vitrification fluid and the same vitrification fluid with open pulled straw(OPS) or glass micro-pipe(GMP).The influence of different priming solution,cryoprotectants,carrier,freezing and thawing methods on developmental capability of bovine oocytes was examined and frozen embryos of bovine also was transferred.1. The GV-oocytes were vitrified by the same vitrification fluid with different concentration adding BAS or FCS. Supplementing concentration of BSA or FCS can influence rates of morphologically normal and matured oocytes.2.The bovine oocytes in different stage(GV and MⅡ) were vitrified. There were no statistical significant differences in rates of morphologically normal (71.3%,68.8%,p>0.05)and rates of matured oocytes(45%,47.6%,p>0.05).There was significant difference in rate of cleavge(23.3%,31.1%,p<0.05).3. Through the contrast impact of vitrification frozen on oocytes with the different number of cumulus cell layer .It was found that,the impact of vitrification frozen on oocyte with 2~4 layers of cumulus cell is better than other number of cumulus cell layer.After vitrification of the oocytes, rates of morphologically normal oocytes were 21.7%,60.3%,60.7%, cleavage rates were 1.6%,30.2%,24.6%.4.In GV-stage oocytes, Through the contrast impact of vitrification on GV-stage oocytes vitrified by the same vitrification fluid with different concentration and different vitrification fluid ,There was no significant difference in rate of morphologically normal oocytes (67.2%,66.7%,p>0.05) and rate of maturation (42.9%,43.7%,p>0.05) with EFS30 or EFS40. But there was significant difference in rate of morphologically normal oocytes and the maturation rate of the oocytes vitrified between EFS and EDFS.5. Through the contrast impact of vitrification frozen on oocyte vitrified by different vitrification fluid and the same vitrification fluid with different concentration, EDFS has better impact of bovine oocyte vitrification frozen.6.In GV-stage oocytes, there was no difference in rates of morphologically normal oocytes(68.4%,74.1%,p>0.05),the maturation rate(47.3%,48.4%,p>0.05)by OPS or GMP with EFS40, while there was difference between EFS40 and EDFS40 vitrification in the maturation rates(48.4%, 49.1%,p<0.05)by GMP .7.The rates of morphologically normal and matured of oocytes with three-step or five-step thawing method were higher than one-step,while there was difference in rates of morphologically normal oocytes(32.3%,51.7%,49.1%,p<0.05),rates of maturation(25.4%,43.4%,36.7%,p<0.05)and rates of cleavage(18.7%,30%,29.8%,p<0.05).8.Effect of transferring bovine ftozen embryos showed that there was no significant differences in pregnancy between one and three-step two methods.Pregnancy rate of bovine embryos frozen by three-step programmed freezing with supplementing concentration of 10% GLY was 38.3%,and pregnantcy rate of bovine embryos frozen by one-step programmed freezing with supplementing concentration of 15% EG was 39.6%.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Cow
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