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培养期间补充白藜芦醇对牛体外生产胚胎质量和耐冻性的影响。

Effect of resveratrol supplementation during culture on the quality and cryotolerance of bovine in vitro produced embryos.

作者信息

Salzano A, Albero G, Zullo G, Neglia G, Abdel-Wahab A, Bifulco G, Zicarelli L, Gasparrini B

机构信息

Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.

Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.

出版信息

Anim Reprod Sci. 2014 Dec 30;151(3-4):91-6. doi: 10.1016/j.anireprosci.2014.09.018. Epub 2014 Oct 2.

Abstract

The aim of the study was to evaluate whether resveratrol supplementation of bovine culture medium improves in vitro blastocyst development, embryo cryotolerance and cell numbers. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, zygotes were cultured in SOF medium, supplemented with 0 (control, n=439), 0.25μM (n=422), 0.5μM (n=447) and 1μM resveratrol (n=416). On Day 7 (IVF=Day 0) blastocysts were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide and 0.5M sucrose. Development rate, i.e. the percentage of embryos resuming development to reach a more advanced stage, and hatching rate were evaluated after 24 and 48h culture. Blastocysts cultured with (0.5μM) and without resveratrol underwent differential staining to count inner cell mass (ICM) and trophectoderm (TE) cells. Resveratrol during culture did not increase blastocyst yields (57.1, 57.7, 59.2 and 46.6%, respectively in 0, 0.25, 0.5 and 1μM resveratrol). However, 0.5μM resveratrol improved embryo cryotolerance compared to the control, as indicated by higher development rates (67.3% vs 50.3%, respectively; P<0.01) and hatching rates (58.9% vs 30.9%, respectively; P<0.01) recorded after 48h post-warming culture. Blastocysts produced in the control and in 0.5μM resveratrol groups had similar numbers of ICM (34.1 and 36.4, respectively), TE (88.1 and 85.3, respectively) and total (122.2 and 121.7, respectively) cells. In conclusion, low levels of resveratrol during in vitro culture improve the quality of IVP bovine embryos, as indicated by their increased resistance to cryopreservation.

摘要

本研究的目的是评估在牛培养基中添加白藜芦醇是否能改善体外囊胚发育、胚胎冷冻耐受性和细胞数量。按照标准程序,将屠宰场采集的卵母细胞进行体外成熟和受精。体外受精20小时后,将受精卵在添加了0(对照,n = 439)、0.25μM(n = 422)、0.5μM(n = 447)和1μM白藜芦醇(n = 416)的SOF培养基中培养。在第7天(体外受精日为第0天),将囊胚用冷冻环在含有16.5%乙二醇、16.5%二甲亚砜和0.5M蔗糖的溶液中进行玻璃化冷冻。在培养24小时和48小时后,评估发育率,即恢复发育至更高级阶段的胚胎百分比,以及孵化率。对添加(0.5μM)和未添加白藜芦醇培养的囊胚进行差异染色,以计数内细胞团(ICM)和滋养外胚层(TE)细胞。培养过程中,白藜芦醇并未提高囊胚产量(在0、0.25、0.5和1μM白藜芦醇组中分别为57.1%、57.7%、59.2%和46.6%)。然而,与对照组相比,0.5μM白藜芦醇提高了胚胎冷冻耐受性,解冻后48小时培养后的发育率(分别为67.3%和50.3%;P<0.01)和孵化率(分别为58.9%和30.9%;P<0.01)更高即表明了这一点。对照组和0.5μM白藜芦醇组产生的囊胚内细胞团数量(分别为34.1和36.4)、滋养外胚层细胞数量(分别为88.1和85.3)和总细胞数量(分别为122.2和121.7)相似。总之,体外培养过程中低水平的白藜芦醇可提高体外生产的牛胚胎质量,这表现为其对冷冻保存的抵抗力增强。

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