培养条件对绵羊卵巢卵母细胞体外成熟及受精的影响

The effects of in vitro culture conditions on maturation and fertilization of ovary oocytes in sheep

对绵羊卵巢卵母细胞体外成熟培养及体外受精培养条件进行筛选,结果表明:在基础培养基中添加FCS可以显著提高卵母细胞的体外成熟率,而添加10%、15%和20%FCS的组间,卵母细胞的成熟率差异不明显,表明在基础培养液中添加10%FCS即可使绵羊卵母细胞的体外成熟率达到一个比较高的水平;体外培养时间不同对绵羊卵母细胞的成熟有较大的影响,培养24 h和26 h的卵母细胞成熟率(分别为73.3%和77.5%)显著高于培养18 h和20 h的卵母细胞成熟率(分别为62.5%和65.0%);对活力较低的绵羊精子而言,采用直接上浮法要比离心洗涤法更有利于选取活力较高的精子,并可提高受精卵的卵裂率.

In the study, 4 068 oocytes from 532 sheep ovaries were used in present studies to investigate the factors that influence the process of the oocyte in vitro maturation（IVM）, in vitro fertilization（IVF） and in vitro culture（IVC） of the early embryos. The results showed that the number of retrieval oocytes per ovary was 4.71,and of total oocytes,2 962 oocytes were level A and B. FCS obviously influenced the rate of IVM. When the concentration of FCS were 10 %, 15% and 20 % in culture medium, the rate of IVM had no significant difference. But compared with FCS free medium group, add FCS can increase the rate of IVM（52 % ,72 % ,respectively）. The rate of IVM can be influenced by culture time,it is higher in group that the oocytes were cultured 24 and 26 hours compared with group that cultured 18 and 20 hours （73.3 % and 77.5 % ,62.5 % and 65.0 % ,respectively ）. For the treatment procedure of sperm,the method of swimming up was more useful than the method of centrifugation and washing, because it can get high viable sperm and high rate of fertilization.