Effect of addition of hyaluronan to embryo culture medium on survival of bovine embryos in vitro following vitrification and establishment of pregnancy after transfer to recipients
Effect of addition of hyaluronan to embryo culture medium on survival of bovine embryos in vitro following vitrification and establishment of pregnancy after transfer to recipients
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DOI：
10.1016/j.theriogenology.2008.11.007
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发表时间：
2009-04-15
期刊：
影响因子：
2.8
通讯作者：
Hansen, P. J.
中科院分区：
文献类型：
--
作者：
Block, J.;Bonilla, L.;Hansen, P. J.
进行了两个实验以确定在培养基中添加乙酰透明质酸是否可以提高玻璃化冷冻后或胚胎移植后牛胚胎的存活率。在实验I中，在体外产生胚胎并在含有四种浓度的乙酰透明质酸（0、0.1、0.5或1mg/mL）之一的改良合成输卵管液（SOF）中培养7天。含或不含 4 mg/mL 牛血清白蛋白 (BSA)。授精后第7天，使用打开的吸管将囊胚和扩张的囊胚玻璃化。浓度为 1 mg/mL。透明质酸增加了 (P < 0.05) 升温后 24 小时的囊胚卵母细胞百分比和再扩张率。在 0.5 mg/mL 浓度下，透明质酸倾向于提高 (P < 0.10) 升温后 48 小时的再膨胀率，并提高 (P < 0.05) 24 小时和 72 小时的胚胎孵化率。 BSA 处理导致卵裂率略有降低 (P < 0.05)，但仅对于含有透明质酸 (BSA X 透明质酸，P = 0.1 0) 的培养物，成为囊胚的卵母细胞百分比增加 (P < 0.001)，并降低再增殖率 (P < 0.001) 和孵化率 (P < 0.05 或 P < 0.01）始终进行检查。在实验2中，在体外产生胚胎并在含有4 mg/mL BSA的改良SOF中培养。含或不含 1 mg/mL 透明质酸。授精后159-162小时，收获1级桑葚胚、囊胚和扩大的囊胚用于胚胎移植。在推定排卵后第 7 天，将收获的胚胎单独移植到具有明显黄体的哺乳期荷斯坦受体中。透明质酸和胚胎阶段对妊娠率存在交互作用（P < 0.05）。接受经过透明质酸处理的桑葚胚和囊胚期胚胎的受者的妊娠率高于接受同一阶段对照胚胎的受者。透明质酸对接受扩大囊胚的受者的妊娠率没有影响。总之，在胚胎培养物中添加透明质酸可提高囊胚产量，提高玻璃化冷冻后的存活率，并提高新鲜桑葚胚和囊胚期胚胎的移植后存活率。 (c) 2009 Elsevier Inc. 保留所有权利。
Two experiments were conducted to determine whether addition of hyaluronan to Culture medium Could improve Survival of bovine embryos after vitrification or following embryo transfer. In Experiment I, embryos were produced in vitro and cultured tor 7 days in modified synthetic oviductal fluid (SOF) containing one Of four concentrations of hyaluronan (0, 0.1, 0.5, or 1 mg/mL). with or without 4 mg/mL of bovine serum albumin (BSA). on Day 7 after insemination, blastocysts and expanded blastocysts were vitrified using open-pulled straws. At a concentration of 1 mg/mL. hyaluronan increased (P < 0.05) the percentage of oocytes that were blastocysts and re-expansion rate at 24 h after warming. At 0.5 mg/mL, hyaluronan tended (P < 0.10) to increase re-expansion rate at 48 h after warming and increased (P < 0.05) embryo hatching rate at 24 and 72 h. Treatment with BSA caused a slight reduction in cleavage rate (P < 0.05), but only for Cultures containing hyaluronan (BSA X hyalauronan, P = 0.1 0) an increase in the percentage of oocytes that became blastocysts (P < 0.001), and a reduction in re-expansion rates (P < 0.001) and hatching rates (P < 0.05 or P < 0.01) at all times examined. In Experiment 2, embryos were produced in vitro and cultured in modified SOF containing 4 mg/mL BSA. with or without 1 mg/mL hyaluronan. At 159-162 h after insemination, grade 1 morula, blastocysts and expanded blastocysts were harvested for embryo transfer. Harvested embryos were transferred individually to lactating Holstein recipients with a palpable corpus luteum on Day 7 after presumptive ovulation. There was an interaction (P < 0.05) between hyaluronan and embryo stage on pregnancy rate. Recipients that received morula and blastocyst stage embryos treated with hyaluronan had a higher pregnancy rate than recipients that received control embryos of the same stage. There was no effect of hyaluronan on pregnancy rates of recipients that received expanded blastocysts. In conclusion, addition of hyaluronan to embryo culture enhanced blastocyst yield, improved survival following vitrification, and enhanced the post-transfer survival of fresh morula and blastocyst stage embryos. (c) 2009 Elsevier Inc. All rights reserved.