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使用新型胚胎培养液生产的新鲜或冷冻胚胎进行泌乳奶牛的体外胚胎移植的效果。

Efficacy of in vitro embryo transfer in lactating dairy cows using fresh or vitrified embryos produced in a novel embryo culture medium.

机构信息

Department of Animal Sciences, University of Florida, Gainesville, FL 32611, USA.

出版信息

J Dairy Sci. 2010 Nov;93(11):5234-42. doi: 10.3168/jds.2010-3443.

Abstract

Objectives were to determine whether pregnancy success could be improved in lactating cows with timed embryo transfer when embryos were produced in vitro using a medium designed to enhance embryo development and survival after cryopreservation. In experiment 1, embryos (n=569 to 922) were cultured in either modified synthetic oviduct fluid or a serum-free medium, Block-Bonilla-Hansen-7 (BBH7). Development to the blastocyst stage was recorded at d 7, and selected blastocysts (n=79 to 114) were vitrified using open pulled straws. Culture of embryos in BBH7 increased development to the blastocyst stage (41.9±2.0 vs. 14.7±2.0%) and advanced blastocyst stages (expanded, hatching, hatched; 31.1±1.3 vs. 6.4±1.3%) at d 7 and resulted in higher hatching rates at 24h postwarming compared with embryos cultured in modified synthetic oviduct fluid (59.0±0.5 vs. 26.7±0.5%). In experiment 2, embryos were produced using X-sorted semen and cultured in BBH7. At d 7 after insemination, embryos were transferred fresh or following vitrification. Lactating Holstein cows were either subjected to timed artificial insemination (TAI) on the day of presumptive ovulation or used as embryo recipients 7 d later. Embryo recipients received an embryo if a corpus luteum was present. The percentage of cows pregnant at d 32, 46, and 76 of gestation was higher among cows that received fresh embryos compared with TAI cows or cows that received vitrified embryos. At d 76, for example, the proportion and percentage pregnant was 47/150 (31.3%) for cows subjected to TAI, 48/95 (50.5%) for cows receiving fresh embryos, and 39/141 (27.7%) for cows receiving a vitrified embryo. No difference was observed in the percentage of cows pregnant among TAI cows and those that received vitrified embryos. There was a service or transfer number × treatment interaction because differences in pregnancy rate between embryo transfer recipients and cows bred by TAI were greater for cows with more than 3 services or transfers. Pregnancy success in lactating cows can be improved by transferring fresh embryos produced in BBH7 compared with TAI. Moreover, no decline in fertility was observed when cryopreserved embryos were transferred compared with TAI. Embryo transfer is particularly efficacious for infertile cows that have previously experienced several failed breeding attempts.

摘要

目的是确定在使用旨在增强胚胎冷冻保存后发育和存活的培养基体外生产胚胎的哺乳期奶牛中,通过定时胚胎移植是否可以提高妊娠成功率。在实验 1 中,将胚胎(n=569 至 922)培养在改良的合成输卵管液或无血清培养基 Block-Bonilla-Hansen-7(BBH7)中。在第 7 天记录胚胎发育至囊胚阶段,并选择囊胚(n=79 至 114)使用开放式拉制吸管进行玻璃化冷冻。胚胎在 BBH7 中的培养增加了囊胚阶段的发育(41.9±2.0%比 14.7±2.0%)和高级囊胚阶段(扩张、孵化、孵化)(31.1±1.3%比 6.4±1.3%)在第 7 天,并导致解冻后 24 小时的孵化率高于在改良的合成输卵管液中培养的胚胎(59.0±0.5%比 26.7±0.5%)。在实验 2 中,使用 X 分选精液生产胚胎,并在 BBH7 中培养。在受精后第 7 天,新鲜胚胎或玻璃化冷冻胚胎进行移植。泌乳荷斯坦奶牛要么在假定排卵日接受定时人工授精(TAI),要么在 7 天后作为胚胎接受者。如果存在黄体,则胚胎接受者会接受胚胎。与 TAI 奶牛或接受玻璃化冷冻胚胎的奶牛相比,接受新鲜胚胎的奶牛在妊娠第 32、46 和 76 天的怀孕百分比更高。例如,在第 76 天,接受 TAI 的奶牛的比例和怀孕百分比为 47/150(31.3%),接受新鲜胚胎的奶牛为 48/95(50.5%),接受玻璃化冷冻胚胎的奶牛为 39/141(27.7%)。接受 TAI 的奶牛和接受玻璃化冷冻胚胎的奶牛之间的怀孕百分比没有差异。在胚胎接受者和通过 TAI 繁殖的奶牛之间的妊娠率差异与服务或转移次数×处理的相互作用有关,因为具有超过 3 次服务或转移的奶牛之间的妊娠率差异更大。与 TAI 相比,用 BBH7 生产的新鲜胚胎进行胚胎移植可以提高哺乳期奶牛的妊娠成功率。此外,与 TAI 相比,冷冻保存胚胎的转移并未观察到生育力下降。胚胎移植对于以前经历过多次繁殖失败的不孕奶牛特别有效。

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