应用1.8mol/L乙二醇+蔗糖冷冻保存奶牛胚胎

Freezing Bovine Embryos in 1 8 mol/L Ethylene Glycol(EG)+Sucrose Solution

以 1 .8 mol/L乙二醇 +0 .1 mol/L蔗糖 +2 0 %FCS-m PBS和 1 .8mol/L乙二醇 +0 .0 5mol/L蔗糖 +2 0 %FCS-m PBS对奶牛胚胎进行了冷冻保存试验 ,并与以 1 .4 mol/L丙三醇 +2 0 %FCS-m PBS常规冷冻保存奶牛胚胎效果进行了比较。冷冻程序为 -7℃平衡 5min后植冰 ,平衡 1 0 min,然后以 0 .3℃ /min的速率降至 -3 3℃ ,直接浸入液氮中保存。胚胎解冻后的培养结果 ,3组之间的胚胎存活率 ,分别为 80 .0 %( 1 6/2 0 )、90 .5%( 1 9/2 1 )、90 .0 %( 1 8/2 0 ) ,差异不显著 ( P>0 .0 5) ;胚胎的囊胚孵化率 ,1 .8mol/L乙二醇 +0 .0 5mol/L蔗糖组达 81 .0 %( 1 7/2 1 ) ,极显著高于 1 .8mol/L乙二醇 +0 .1 mol/L蔗糖组的 3 0 .0 %( 6/2 0 )和 1 .4mol/L丙三醇组的 2 5.0 %( 5/2 0 ) ( P<0 .0 1 )。 1 .8mol/L乙二醇 +0 .0 5mol/L蔗糖组冷冻的 A级胚胎和 B级胚胎之间的存活率差异不显著 ( P>0 .0 5) ,但是 A级胚胎的囊胚孵化率 ( 1 0 0 %,1 1 /1 1 )显著高于 B级胚胎( 60 %,6/1 0 ) ( P<0 .0 5)。另外 ,胚胎解冻后透明带断裂对其发育率无影响 ;冷冻前胚胎在 1 .8mol/L乙二醇+0 .0 5mol/L蔗糖液中平衡 1 0 min和平衡 2 0 min,冷冻解冻后培养发育率差异不显著 ( P>0 .0 5)。试验结果表明 ,应用 1 .8mol/L乙二醇 +0 .

Embryos were divided into three groups Embryos in group 1,group 2,and group 3 were frozen in 1 8 mol/L EG+0 1 mol/L sucrose+20% FCS(fetal cow serum) mPBS,1.8 mol/L EG+ 0.05 mol/L sucrose+20% FCS mPBS and 1.4 mol/L glyceral+20% FCS mPBS solutions respectively They were held at -7 ℃ for 5 minutes,seeded,and held again for 10 minutes,cooled again at -0 3 ℃/minute to -33 ℃,then plunged and stored in liquid nitrogen The survival rates of the three groups were 80 0%(16/20),90 5%(19/21),90 0(18/20) respectively,there were no significantly differences among them (P>0 05) The rate of embryos that developed into hatched blastocysts in group 2(81 0%,17/21) was significantly higher than that in group 1(30 0%,6/20) and in group 3(25%,5/20)(P<0 01) The survival rate of excellent embryos in group 2 was not significantly different from that of good embryos in group 2(P>0 05),but the rate of excellent embryos that developed into hatched blastocyst(100%,11/11) was significantly higher than that of good embryos(60 0%,6/10) (P<0 05). Furthermore,the cleavage of zona pellucida didn′t affect the developmental rate of the thawed embryos After being cultured,the developmental rate of thawed embryos that had been equiponderated in 1 8 mol/L EG+0 05 mol/L sucrose solution for 10 minutes wasn′t different from that for 20 minutes(P>0 05) The results of this experiment showed that a higher survival rate and hatched rate would be obtained by frozen embryos in 1 8 mol/L EG+0 05 mol/L sucrose solution