Effect of estrous cow serum during bovine embryo culture on blastocyst development and cryotolerance after slow freezing or vitrification
Effect of estrous cow serum during bovine embryo culture on blastocyst development and cryotolerance after slow freezing or vitrification
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DOI：
10.1016/j.theriogenology.2005.08.020
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发表时间：
2006-05-01
期刊：
影响因子：
2.8
通讯作者：
Alberio, RH
中科院分区：
文献类型：
--
作者：
Mucci, N;Aller, J;Alberio, RH
本研究探讨了牛胚胎培养过程中添加发情牛血清（ECS）对囊胚发育和玻璃化冷冻后囊胚存活的影响。胚胎来源于阿巴托衍生的卵母细胞的体外成熟（IVM）和体外受精（IVF）。在第3天，在三种不同的培养基中培养胚胎：Charles Ronsenkrans培养基+氨基酸（CRIaa;不含牛血清白蛋白（BSA））+ 5%发情期牛血清（CR 1-ECS）、CR 1aa + 3 mg/mL BSA（CR 1-BSA）或CR 1aa + 5% ECS + 3 mg/ mL BSA（CR 1-ECS-BSA）。在授精后7.5 d（PI），评价囊胚产量和质量;通过开放式吸管（OPS）玻璃化法或缓慢冷冻（1.5 M乙二醇，EM）冷冻保存来自每种培养基的囊胚和扩张囊胚。CR 1-ECS、CR 1-BSA和CR 1-ECS-BSA的总囊胚率差异无显著性（分别为30.9、33.1和32.9%，P < 0.05）。玻璃化冷冻胚胎的存活率（孵化率）高于慢速冷冻胚胎（分别为43%和12%，P < 0.01）; CR 1-BSA培养胚胎的存活率（40.3%）高于含血清培养胚胎的存活率（CR 1-ECS，21.5%和CR 1-ECS-BSA，19.8%; P < 0.01）。最后：（a）在无血清培养基中体外生产牛胚胎是可能的，而不影响胚泡产量和质量;（B）无血清培养基生产的胚胎质量最好（就冷冻保存后存活率而言）;（c）玻璃化冷冻产生最高的冷冻保存后存活率，无论培养基中是否存在血清。(C)2005年爱思唯尔公司All rights reserved.
The present Study investigated the effect of estrous cow serum (ECS) during culture of bovine embryos on blastocyst development and survival after cryopreservation by slow freezing or vitrification. Embryos were derived from in vitro maturation (IVM) and in vitro fertilization (IVF) of abbatoir-derived oocytes. At Day 3, embryos were Cultured in three different media: Charles Ronsenkrans medium + amino acids (CRIaa; without bovine serum albumin (BSA)) + 5% estrous cow serum (CR1-ECS), CR1aa + 3 mg/mL BSA (CR1-BSA) or CR1aa + 5% ECS + 3 mg/ mL BSA (CR1-ECS-BSA). At 7.5 d post-insemination (PI), blastocyst yield and quality were evaluated; blastocysts and expanded blastocysts from each media were cryopreserved by Open Pulled Straw (OPS) vitrification method or slow freezing (1.5 M ethylene glycol, EM). Total blastocyst yield did not differ among CR1-ECS, CR1-BSA and CR1-ECS-BSA (30.9, 33.1 and 32.9%, respectively, P < 0.05). Embryo Survival (hatching rate) was higher in vitrified Versus slow-frozen embryos (43% versus 12%, respectively, P < 0.01), and in embryos cultured in CR1-BSA (40.3%) compared with those cultured in serum-containing media (CR1-ECS, 21.5% and CR1-ECS-BSA, 19.8%; P < 0.01). In conclusion: (a) it was possible to produce in vitro bovine embryos in serum-free culture medium without affecting blastocyst yield and quality; (b) serum-free medium produced the best quality embryos (in terms of post-cryopreservation survival); and (c) vitrification yielded the highest post-cryopreservation survival rates, regardless of the presence of serum in the culture medium. (C) 2005 Elsevier Inc. All rights reserved.