秦川牛胚胎冷冻保存效果

Qinchuan Cattle Embryo Cryopreservation

为探明秦川牛胚胎冷冻保存技术的可行性,将48头秦川牛母牛进行多次超数排卵处理、人工授精,并于授精后第7天非手术法获得胚胎,把获得的A级胚用不同浓度甘油(1.0~5.0 mol/L)和乙二醇(1.0~5.0mol/L)进行处理,并采用常规冷冻法和快速冷冻法进行冷冻,解冻后进行体外培养和不同发育阶段胚胎(桑椹胚、囊胚和扩张囊胚)移植,统计其存活率、孵化率和妊娠率等。结果表明:秦川牛7日龄胚胎能耐受甘油的浓度为2.0~4.0mol/L,乙二醇为1.0~3.0mol/L;常规冷冻法解冻后胚胎的存活率和孵化率显著(P<0.05)高于快速冷冻法,胚胎移植妊娠率和产犊率差异不显著(P>0.05);常规冷冻桑椹胚的冻后存活率、孵化率极显著(P<0.01)低于囊胚和扩张囊胚,移植妊娠率和产犊率显著(P<0.05)低于囊胚和扩张囊胚。

In order to investigate the feasibility of Qinchuan cattle embryo cryopreservation technology,48 female Qinchuan cattle were super-ovulated repeatedly,then artificial inseminated,and the embryos were obtained non-surgically in 7th day after artificial insemination.The A-class embryos were treated with different concentrations of glycerol(1.0~5.0 mol/L)and ethylene glycol(1.0~5.0 mol/L),and frozen by conventional method and rapid method.After thawed,in vitro cultured and transplanted the embryos in different developmental stages(morulas,blastocysts and expanded blastocysts).Results:7-day old embryo of Qinchuan cattle can tolerate glycerol concentration range of 2.0~4.0 mol/L,ethylene glycol 1.0~3.0mol/L.The survival rate and hatching rate of conventional freezing method was significantly(P<0.05)higher than that of rapid freezing method;The morula's survival rate and hatching rate after frozen and thawed was very significantly(P<0.01)lower than that of blastocyst and expanded blastocyst;Pregnancy rate and calving rate of embryos frozen by conventional method and rapid method had no significant(P>0.05)difference after transplantation.The freezing morula's pregnancy rate and calving rate was significantly(P <0.05)lower than that of blastocyst and expanded blastocyst after transplantation.