和牛OPU卵母细胞体外受精后胚胎发育潜能研究  

Study on Embryonic Developmental Potential of Embryos Derived from Ovum Pickup Oocytes and Fertilization in vitro in Wagyu

作　　者：徐光勇 刘振田 陈祥飞 张美洁 杜涛 张俊波 刘智辉 邢雪松 杜福良[2] XU Guangyong;LIU Zhentian;CHEN Xiangfei;ZHANG Meijie;DU Tao;ZHANG Junbo;LIU Zhihui;XING Xuesong;DU Fuliang(Qingdao Longmingniu Embryo Biotechnologies Co.,Ltd.,Qingdao 266400,China;College of Life Sciences,Nanjing Normal University,Nanjing 210023,China)

机构地区：[1]青岛隆铭牛胚胎生物技术有限公司,青岛266400 [2]南京师范大学生命科学学院,南京210023

出　　处：《中国畜牧兽医》2025年第2期730-739,共10页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金委面上项目(32372877)。

摘　　要：[目的]在体外成熟液(IVM)中添加不同来源的卵泡刺激素(FSH)和黄体生成素(LH)对日本黑牛(和牛)活体采卵(OPU)卵母细胞进行成熟培养,研究其体外受精(IVF)的胚胎在体外培养(IVC)与体内发育的潜能。[方法]按照3×2试验设计,OPU卵母细胞在3种FSH(NIH-FSH、NB-FSH和CA-FSH,0.13 IU/mL)和2种LH(NIH-LH和NB-LH,23 IU/mL)匹配的6组IVM液中培养22~24 h, IVF和IVC后检查囊胚发育率。将胚胎移植(ET)至同步化受体后40 d,采用超声波检查妊娠率。[结果]最佳匹配组是在IVM培养液中添加0.13 IU/mL NIH-FSH与23 IU/mL NIH-LH,囊胚发育率最高达47.3%。OPU卵母细胞经IVM和IVF后,在体外培养6 d的胚胎(IVC 6D)妊娠率(62.6%~64.0%)与体内胚胎相比无显著差异(P>0.05),但显著高于体外培养7 d的胚胎(IVC 7D)(44.2%~47.1%)(P<0.05)。囊胚移植到青年牛受体的妊娠率高达67.1%,移植后未孕的受体可继续ET,但ET重复4次时妊娠率显著低于0、1次(P<0.05)。[结论]不同来源的FSH与LH的IVM匹配对OPU-IVF的胚胎发育潜能有明显的影响。移植IVC 6D的囊胚比IVC 7D的妊娠率显著升高。本研究优化的OPU-IVF-ET体系可有效培育优良的和牛品系,并快速扩繁其生产群体。[Objective]Oocytes collected by ovum pickup(OPU)in Japanese Black cattle(Wagyu)were matured in vitro(IVM)in medium supplemented with different sources of follicle-stimulating hormone(FSH)and luteinizing hormone(LH),and the in vitro and in vivo developmental potentials were studied after in vitro fertilization(IVF)and in vitro culture(IVC).[Method]According to the 3×2 experimental design,OPU oocytes were matured in 6 groups of IVM media in combinations with 3 FSH(NIH-FSH,NB-FSH and CA-FSH,0.13 IU/mL)and 2 LH(NIH-LH and NB-LH,23 IU/mL)for 22-24 h.The blastocyst developmental rate was compared after IVF and IVC.The pregnancy rate was examined by ultrasound 40 days after embryo transfer(ET)to the synchronized recipients.[Result]The best combination was supplementing 0.13 IU/mL NIH-FSH and 23 IU/mL NIH-LH in IVM medium,and the blastocyst development rate was up to 47.3%.The pregnancy rate of blastocysts(62.6%-64.0%)derived from OPU oocytes cultured for 6 days(IVC 6D)post IVM and IVF was not significantly different from that of in vivo embryos(P>0.05),but significantly higher than that of embryos cultured for 7 days(IVC 7D)(44.2%-47.1%)(P<0.05).The pregnancy rate of blastocysts transferred into heifer recipients was as high as 67.1%,and open recipients after pervious ET cycle could be used for next ET treatment,but the pregnancy rate was significantly lower when ET was repeated 4 times compared with 0 and 1 times(P<0.05).[Conclusion]This study demonstrated that different sources of FSH and LH in combinations in IVM medium had a significant effect on the embryonic development potential of OPU-IVF oocytes.The pregnancy rate of blastocysts from IVC 6D was significantly higher than that from IVC 7D.The optimized OPU-IVF-ET system could effectively cultivate excellent Wagyu breed and rapidly expand its production population.

关 键 词：和牛 活体采卵 体外受精 胚胎移植 妊娠 

分 类 号：S814.8[农业科学—畜牧学]