玻璃化冷冻山羊卵母细胞效果研究

Study on the effect of vitrification on goat oocytes

为了提高玻璃化冷冻保存山羊卵母细胞的效果,试验用不含Ca2+的玻璃化冷冻液1[30%乙二醇(EG)]、玻璃化冷冻液2[27.5%EG+2.5%二甲基亚砜(DMSO)]和玻璃化冷冻液3(25%EG+5%DMSO)研究抗冻保护剂DMSO和EG的不同浓度配比对山羊卵母细胞形态正常率、孤雌激活率、体外受精卵裂率及胚胎发育能力的影响。结果表明:山羊卵母细胞经玻璃化冷冻液1,2,3处理后细胞形态正常率(97.1%、97.3%、97.3%)与对照组(100%)相比有所降低,但差异不显著(P>0.05),且对卵母细胞均不产生具有化学毒性作用的孤雌激活;山羊卵母细胞经3种玻璃化冷冻液冷冻保存后,玻璃化冷冻液2冷冻保存卵母细胞的囊胚率(13.8%)显著高于玻璃化冷冻液1(5.7%)和玻璃化冷冻液3(5.1%)(P<0.05)。说明采用玻璃化冷冻液2冷冻保存山羊卵母细胞可获得较为理想的效果。

To improve the effect of vitrification cryopreservation on goat oocytes, this experiment was conducted to use three vitrification solutions （VS） without Ca2＋, containingVSl（30% EG）, VS2 （27.5%EG＋2.5% DMSO） and VS3 （25%EG＋5% DMSO）, to investigate the effects of different concentrations of DMSO and EG as antifreeze agents on the nomml morphology rate, parthenogenetic activation rate, in vitro fertilization cleavage rate and embryonic development in goat oocytes. The results showed that the normal morphology rates of oocytes treated with the three vitrification solutions, were 97.1%, 97.3% and 97.3%, respeetively. There were no significant decreases （ P 〉 0. 05） in the normal morphology rates compared to the control group（ 100% ）, and no parthenogenetic activation with toxic effects was observed in the ooeytes treated with different vitrification solutions. After the goat oocytes were cryopreserved with the three vitrification solutions, the blastocyst rate of the oocytes cryopreserved with the VS2 was higher （ P 〈0.05 ） than that cryopreserved with the VS1 and VS3. It is concluded that eryopreserva- tion of goat oocytes with the VS2（27.5% EG ＋ 5% DMSO） ean obtain more desirable effect.