和牛OPU卵母细胞体外受精后胚胎发育潜能研究

doi:10.16431/j.cnki.1671-7236.2025.02.023

摘要/Abstract

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。

关键词: 和牛; 活体采卵; 体外受精; 胚胎移植; 妊娠

Abstract: 【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.

Key words: Wagyu; ovum pickup; in vitro fertilization; embryo transfer; pregnancy

中图分类号:

S814.8

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. 和牛OPU卵母细胞体外受精后胚胎发育潜能研究[J]. 中国畜牧兽医, 2025, 52(2): 730-739.

XU Guangyong, LIU Zhentian, CHEN Xiangfei, ZHANG Meijie, DU Tao, ZHANG Junbo, LIU Zhihui, XING Xuesong, DU Fuliang. Study on Embryonic Developmental Potential of Embryos Derived from Ovum Pickup Oocytes and Fertilization in vitro in Wagyu[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 730-739.

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