甲基-β-环化糊精对猪体外受精及早期胚胎发育的影响

为了优化猪体外受精技术体系,本试验探索了甲基-β-环化糊精(methyl-beta-cyclic dextrin,MBCD)对猪体外受精以及早期胚胎发育的影响。在体外受精0和4 h向受精液(modified Tris-buffered medium,mTBM)中添加不同浓度(0,0.5,1,2,5,10,15,20μmol/mL)的MBCD,受精孵育结束后转至PZM-3培养液中进行胚胎培养。对各处理组卵母细胞的受精情况以及胚胎发育能力进行了系统的检测,并用金霉素(chlortetracycline,CTC)染色法评估了MBCD处理后精子获能状态。结果显示:1)体外受精0 h添加5μmol/mL MBCD组的卵裂率、囊胚率、囊胚细胞数显著高于(P<0.05)对照组和除10μmol/mL MBCD组之外的其他试验组。2)体外受精0 h添加5和10μmol/mL MBCD组、单精入卵率显著高于(P<0.05)对照组和其他试验组,而多精入卵率显著低于(P<0.05)对照组和其他试验组。3)添加5μmol/mL MBCD组,0～1 h,F型精子迅速减少(78.56～19.43),B型精子迅速增加(10.79～69.86);1～4 h,F型精子和B型精子基本保持不变(B型:69.86～78.78,F型:19.43～9.11)。上述结果表明在体外受精0 h向mTBM中加入5μmol/mL MBCD可以显著提高获能精子比例,减少多精受精发生,提高早期胚胎发育潜能。

Effect of methyl-beta-cyclicdextrin (MBCD) on porcine in vitro fertilization and early embryo development

In order to optimize the porcine IVF technology system,this experiment was designed to explore the influence of methyl-beta-cyclic dextrin(MBCD)on porcine in vitro fertilization and early embryo development.Different concentrations(0,0.5,1,2,5,10,15,20 μmol/mL) of MBCD was added to modified Tris-buffered medium(mTBM) at 0 h and 4 h of in vitro fertilization then the resulted embryos were cultured in PZMμ3.Embryo in vitro development rate and polyspermy proportion was examined,and sperm capacitation state was assessed by chlortetracycline(CTC) staining.Results:(1)In the group of adding 5 μmol/mL MBCD to mTBM at IVF 0 h the cleavage rate,blastocyst rate,cell number of blastocyst was significantly(P<0.05) higher than that in the control group and the other experimental group except 10 μmol/mL group;(2)In the group of 5 and 10 μmol/mL MBCD to mTBM at IVF 0 h the monospermy rate was significantly(P<0.05) higher than that in the control group and the other experimental groups,and the polyspermy rate in group of 5 and 10 μmol/mL MBCD was significantly(P<0.05) lower than that in the control group and the other experimental groups;(3)In the group of 5 μmol/mL MBCD to mTBM at IVF 0 h,during 0-1 h of incubation,the number of F type sperm rapidly reduced(78.56-19.43);while the number of B type sperm increased rapidly(10.79-69.86).During 1-4 h of the incubation,the number of F type and B type sperm had not much change(B: 69.86-78.78,F: 19.43-9.11).In conclusion,the addition of 5 μmol/mL MBCD at 0 h of IVF can significantly improve capacitation sperm percentage,reduce polyspermy fertilization,and thereby enhance the early embryo development competence.