【摘要】 旨在探讨牦牛和黄牛种间体外受精和胚胎发育效果。通过研究黄牛和牦牛卵母细胞体外成熟时间、卵母细胞质量、精子处理方式、受精卵培养体系及冷冻—解冻后胚胎活力。结果表明:随着体外培养时间的延长,卵母细胞第一极体排出率增加,A级卵母细胞培养24h第一极体的排出率显著高于B级和C级卵母细胞,但相同体外成熟时间和相同分级的黄牛和牦牛卵母细胞第一极体的排出率差异不显著。用Percoll液梯度离心法和BO液洗涤法分别处理牦牛精液和黄牛精液后通过种间体外受精,BO液洗涤法处理的精液受精后的卵裂率显著高于Per-coll液梯度离心法处理精液受精后的卵裂率,而2种方法处理精液受精后其囊胚率差异不显著。2种方法(黄牛♀×牦牛♂vs.牦牛♀×黄牛♂)生产的早期犏牛胚胎用输卵管上皮细胞共培养和卵丘细胞共培养其囊胚率和孵化囊胚率都显著高于SOF液培养,而3种培养方法培养的早期犏牛胚胎的卵裂率差异不显著。2种IVF胚胎冷冻解冻后形态正常胚(90.52%±5.94%vs.91.53%±7.34%)、24h的存活率(61.45%±8.44%vs.64.72%±11.82%)和48h的存活率(44.81%±6.21%vs.48.47%±10.04%)均差异不显著。将黄牛♀×牦牛♂IVF犏牛胚胎移植到自然发情4头受体黄牛和3头受体犏牛,4头牛怀孕并产犊,妊娠率为57.14%,平均妊娠期258d。更多还原

【Abstract】 The aim was to explore the efficience of in vitro fertilization and embryo development of the cattle-yak.The experiment was conducted to investigate the effects of in vitro maturation time,oocyte grades,different treatment methods of the semen,in vitro culture system and embryonic vitality and development after freezing and thawing on in vitro fertilization cattle×yak.The results showed that following the prolonging of in vitro maturation time,the extruding of the first polar body(Pb1) increased.And the Pb1 of grade A oocytes cultured in 24 h was significantly higher than that of B and C grade oocytes.But there was no significant difference between cattle oocytes and yak oocytes in same culture time and grade.Cleavage rate with BO fluid washing centrifugal process group is significantly higher than that of the Percoll group,but no significant differences on the blastocyst rate.Two kings of IVF cattle-yak embryos(cattle♀×yak♂vs.yak♀×cattle♂) co-culture with oviductal epithelial cells,cumulus cells and SOF medium and there was no significant difference in cleavage rate,but blastocyst rate and hatched blastocyst rate of co-cultured group were significantly higher than that of SOF medium group.The rate of normal morphology embryo(90.52%±5.94% vs.91.53%±7.34%),survival rate within 24 h(61.45%±8.44% vs.64.72%±11.82%) or 48 h(44.81%±6.21% vs.48.47%±10.04%) of the embryos after thawing were no significant difference.Taking 7 bovines including 4 yellow cattle and 3 cattle×yak as recipients,which were transplanted after natural estrus.The pregnancy rate was 57.14% and the length of average pregnancy was 258 days.更多还原