Gou Ke-Mian, Guan Hong, Bai Jia-Hua, Cui Xiu-Hong, Wu Zhi-Fang, Yan Feng-Xiang, An Xiao-Rong
The State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
Anim Reprod Sci. 2009 Jun;112(3-4):316-24. doi: 10.1016/j.anireprosci.2008.05.008. Epub 2008 May 15.
The practicality of using juvenile in vitro embryo transfer (JIVET) on a field scale in China was evaluated in each of three seasons (summer, autumn and winter) from 2006 to 2007. A total of 102 donor Merino lambs (18 summer, 69 autumn and 15 winter) aged 4-8 weeks were stimulated with 4 x 40 mg FSH administered at 12h intervals plus 400 IU PMSG given at the time of the first FSH treatment. Overall, 89.2% (91/102) of the lambs exhibited follicle development and 79.1+/-65.5 (mean+/-S.D.) cumulus-oocyte complexes were recovered per donor lamb. Compared with the groups of summer (84.9+/-55.3) and autumn (83.6+/-70.8) lambs, the number of recovered cumulus-oocyte complexes was significantly decreased in winter (51.4+/-43.7; p<0.05). After recovery, the cumulus-oocyte complexes were matured and fertilized in vitro using frozen-thawed semen and culture in synthetic oviduct fluid medium to the 2-4-c stage of development, when they were transferred surgically in groups of 3-8 (5.33+/-1.47) to the ipsilateral uterine horn of a total of 603 synchronized recipients. The overall mean proportion of cumulus-oocyte complexes developing to 2-c embryos was 61.4% (4308/7013) and differed significantly between seasons (summer 38.5%, autumn 66.1%, winter 74.6%; p<0.01). Pregnancy rate assessed by ultrasound examination approximately 60 days after embryo transfer was 54.4% (328/603) overall, and 36.7% (221/603) of the recipients maintained their pregnancy to full-term, producing an average 1.49 (330/221) offspring, of which 1.21 (267/221) were viable and healthy lambs, per pregnant recipient. Pregnancy rate at day 60 was affected by season (summer 40.5%, autumn 56.7%, winter 55.7%; p<0.05), but did not differ significantly between seasons at full-term (summer 34.2%, autumn 38.9%, winter 30.4%; p>0.05). Based on the number of donors stimulated, the total number of offspring and viable progeny produced per donor lamb in autumn (5.81 and 4.87) was significantly (p<0.01) higher than that of summer (2.79 and 1.94) and winter (4.24 and 3.31). This study showed that each donor lamb after stimulation produced an average of 48.6 transferable embryos that resulted in 4.04 viable and healthy progeny. These results indicate that JIVET is a cost-effective method of multiplying desirable sheep genotypes in China.
2006年至2007年的三个季节(夏季、秋季和冬季),对在中国田间规模使用幼年体外胚胎移植(JIVET)的实用性进行了评估。总共102只4至8周龄的供体美利奴羔羊(夏季18只、秋季69只、冬季15只),每隔12小时注射4×40毫克促卵泡素(FSH)进行刺激,并在首次FSH处理时给予400国际单位孕马血清促性腺激素(PMSG)。总体而言,89.2%(91/102)的羔羊出现卵泡发育,每只供体羔羊回收的卵丘-卵母细胞复合体(COCs)数量为79.1±65.5(平均值±标准差)。与夏季(84.9±55.3)和秋季(83.6±70.8)羔羊组相比,冬季回收的卵丘-卵母细胞复合体数量显著减少(51.4±43.7;p<0.05)。回收后,卵丘-卵母细胞复合体在体外使用冻融精液进行成熟和受精,并在合成输卵管液培养基中培养至2-4细胞发育阶段,然后以3-8个(5.33±1.47)一组通过手术移植到总共603只同期受体的同侧子宫角。发育到2细胞胚胎的卵丘-卵母细胞复合体的总体平均比例为61.4%(4308/7013),且在不同季节间差异显著(夏季38.5%,秋季66.1%,冬季74.6%;p<0.01)。胚胎移植后约60天通过超声检查评估的妊娠率总体为54.4%(328/603),36.7%(221/603)的受体维持妊娠至足月,每只妊娠受体平均产仔1.49只(330/221),其中1.21只(267/221)为存活且健康的羔羊。60天时的妊娠率受季节影响(夏季40.5%,秋季56.7%,冬季55.7%;p<0.05),但足月时不同季节间无显著差异(夏季34.2%,秋季38.9%,冬季30.4%;p>0.05)。基于刺激的供体数量,秋季每只供体羔羊产生的后代总数和存活后代数量(5.81和4.87)显著高于夏季(2.79和1.94)和冬季(4.24和3.31)(p<0.01)。本研究表明,每只受刺激的供体羔羊平均产生48.6个可移植胚胎,可产出4.04只存活且健康的后代。这些结果表明,幼年体外胚胎移植是在中国繁殖优良绵羊基因型的一种经济有效的方法。