猪卵母细胞在玻璃化冷冻液中的培养效果

本试验的目的是要选择较好的用于猪卵母细胞玻璃化冷冻的玻璃化冷冻液，玻璃化液由ＴＣＭ－１９９、ｍＰＢＳ、Ｋｉｅｖ液分别加２０％的犊牛血清与３种抗冷冻剂（丙三醇、１，２──丙二醇和乙二醇）组成。猪卵母细胞采自屠宰厂的卵巢并培养在ＴＣＭ－１９９＋２０％犊牛血清、ｍＰＢＳ＋２０％犊牛血清和Ｋｉｅｖ＋２０％犊牛血清中，在３７℃、湿度１００％条件下培养至少５个小时。５小时后逐渐加入抗冷冻剂，使其终浓度达５０％。培养５分钟后，将猪卵母细胞依次移至抗冷冻剂浓度为２５％，１２．５％和０％的培养液中，培养５分钟，而后进行台盼蓝死活染色。结果表明猪卵母细胞在各组培养后的存活率没有显著性差异（Ｐ＞０．０５），但乙二醇结果优于其它抗冷冻剂，而ＴＣＭ－１９９好于其它两类培养液。

Effect of Vitrificational Solution on Porcine Oocyte

The aim of the test was to deal with different vitrificational solutions with glycol or glycerol or 1,2- propylene glycol in the porcine oocyte. After preincubation for 5 hours in 37℃, 100% air moist in the TCM199 or mPBS or Kiev with Fetal Calf Serum (FCS) respectively. Each of three antifreeze constitu- tents added to culture fluid gradually till 60% concentration. Five minutes later , Porcine ocoytes moved tothesame fluid containing 0%, 12.5%and 25% one of antifreeze constitutents respectively and for five minutes treatment followed by staining with trypan blue. The results indicated the motility ration of porcine oocyte had no difference (P>0. 05 ) in all kinds of fluids. Solutions with ethanol were better than other ones and incubation of porcine oocyte in TCM 199 led to best result in our test.