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CSF2 和 DKK1 对牛胚胎发育和妊娠结局的作用受胚胎培养液成分的影响。

Actions of CSF2 and DKK1 on bovine embryo development and pregnancy outcomes are affected by composition of embryo culture medium.

机构信息

Department of Animal Sciences, D.H. Barron Reproductive and Perinatal Research Program, and Genetics Institute, University of Florida, Gainesville, FL, 32611-0910, USA.

Zoetis, Kalamazoo, MI, 49007, USA.

出版信息

Sci Rep. 2022 May 7;12(1):7503. doi: 10.1038/s41598-022-11447-7.

Abstract

Procedures for in vitro embryo production in cattle have not been optimized. In the current experiment, we utilized a 3 × 3 factorial design to test whether the proportion of embryos becoming blastocysts in culture and the pregnancy rate after embryo transfer are affected by type of serum in the medium [no serum; 3% (v/v) KnockOut Serum Replacement (SR); 3% (v/v) fetal bovine serum (FBS)] and addition of specific embryokines [vehicle; 10 ng/mL colony stimulating factor 2 (CSF2); 100 ng/mL dickkopf related protein 1 (DKK1)] at day 5 of culture. Embryos were produced using abattoir-derived ovaries and Y-sorted semen from two Angus sires. The percent of putative zygotes and cleaved embryos becoming blastocysts was improved by SR and FBS. Pregnancy rate at day 30 was determined for 1426 Nelore recipients and calving rate for 266 recipients. In the absence of CSF2 or DKK1, pregnancy rates were lower for embryos cultured with SR or FBS. CSF2 and DKK1 reduced pregnancy rate for embryos cultured without serum but had no detrimental effect in the SR or FBS groups. Indeed, CSF2 blocked the negative effect of FBS on pregnancy rate. Data on birth weights were available for 67 bull calves. There were no effects of treatment. The sire used to produce embryos had significant and large effects on development to the blastocyst stage, pregnancy rate at day 30, calving rate and pregnancy loss between day 30 and calving. Results indicate that (1) SR and FBS can improve embryonic development in vitro while also compromising competence of embryos to survive after transfer, (2) actions of CSF2 and DKK1 depend upon other characteristics of the embryo production system, and (3) sire can have a large effect on embryonic development before and after transfer.

摘要

牛的体外胚胎生产程序尚未优化。在本实验中,我们利用 3×3 析因设计来测试胚胎在培养中成为囊胚的比例和胚胎移植后的妊娠率是否受到培养基中血清类型[无血清;3%(v/v)无血清替代物(SR);3%(v/v)胎牛血清(FBS)]和添加特定胚胎因子[载体;10ng/mL 集落刺激因子 2(CSF2);100ng/mL 抑制素相关蛋白 1(DKK1)]的影响,添加时间为培养第 5 天。胚胎是使用屠宰场来源的卵巢和来自两个安格斯种公牛的 Y 分选精液生产的。SR 和 FBS 提高了假定受精卵和分裂胚胎成为囊胚的比例。对 1426 头尼洛雷牛受体进行了 30 天妊娠率的测定,对 266 头受体进行了产犊率的测定。在没有 CSF2 或 DKK1 的情况下,用 SR 或 FBS 培养的胚胎妊娠率较低。CSF2 和 DKK1 降低了无血清培养胚胎的妊娠率,但在 SR 或 FBS 组中没有不良影响。事实上,CSF2 阻断了 FBS 对妊娠率的负面影响。67 头公牛犊的出生体重数据可用。处理没有影响。用于生产胚胎的公牛对胚胎发育到囊胚阶段、30 天妊娠率、产犊率和 30 天至产犊期间的妊娠损失有显著的和大的影响。结果表明:(1)SR 和 FBS 可以提高胚胎的体外发育,但也会降低胚胎移植后的生存能力;(2)CSF2 和 DKK1 的作用取决于胚胎生产系统的其他特征;(3)公牛对胚胎的发育有很大的影响,无论是在移植前还是移植后。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4700/9079070/a34d01feb97e/41598_2022_11447_Fig1_HTML.jpg

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