Co-culture with granulosa cells improve the in vitro maturation ability of porcine immature oocytes vitrified with cryolock
Co-culture with granulosa cells improve the in vitro maturation ability of porcine immature oocytes vitrified with cryolock
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DOI：
10.1016/j.cryobiol.2014.08.004
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发表时间：
2014-10-01
期刊：
影响因子：
2.7
通讯作者：
Betancourt, Miguel
中科院分区：
文献类型：
--
作者：
Casillas, Fahiel;Teteltitla-Silvestre, Mario;Betancourt, Miguel
本研究旨在评估两种不同的设备：超细开放牵拉吸管（SOPS）和Cryolock的两种卵母细胞玻璃化-加温程序的效率，以及玻璃化冷冻的未成熟卵母细胞与新鲜颗粒细胞共培养对提高体外成熟（IVM）的影响。用两种程序玻璃化冷冻未成熟卵母细胞：A）将卵母细胞暴露于浓度从4%增加至35%的乙二醇（EG）和0.5M海藻糖。然后，将其装入SOPS或Cryolock中。为了加温，将卵母细胞暴露于浓度递减的海藻糖0.3、0.2和0.1 M中进行IVM。B）将卵母细胞暴露于EG和二甲亚砜（Me 2SO）的两种混合物，7.5%和16%，均含有0.4 M蔗糖，然后加载到SOP或Cryolock中并储存在液氮中。为了温热，将卵母细胞暴露于单一浓度0.5 M的蔗糖中。升温后，测定活力; IVM 44小时后，评价活力和减数分裂阶段。结果表明，在所有成熟阶段，采用SOPS的程序A和B之间没有显著差异，达到21%的最大成熟率。对于Cryolock，在两种程序之间观察到显著差异，程序B更有效，在MII期的产率为38%，并且由于与新鲜颗粒细胞共培养而增加到49%。总之，在玻璃化冷冻的未成熟卵母细胞中，Cryolock和程序B与共培养系统提高了存活率和成熟率。(C)2014爱思唯尔公司All rights reserved.
This study was designed to evaluate the efficiency of two oocyte vitrification-warming procedures using two different devices: Superfine Open Pulled Straws (SOPS) and Cryolock, as well as the effect of the co-culture of vitrified immature oocytes with fresh granulosa cells to improve in vitro maturation (IVM). Immature oocytes were vitrified with two procedures: A) Oocytes were exposed to an increasing concentration of ethylene glycol (EG) from 4% to 35% with 0.5 M trehalose. They then, were loaded in SOPS or Cryolock. For warming, oocytes were exposed to decreasing concentrations of trehalose 0.3, 0.2 and 0.1 M for IVM. B) Oocytes were exposed to two mixtures of EG and dimethylsulfoxide (Me2SO), at 7.5% and 16%, both with 0.4 M of sucrose and then loaded in SOPS or Cryolock and stored in liquid nitrogen. For warming, oocytes were exposed to a single concentration of sucrose 0.5 M. After warming, viability was determined; and after 44 h of IVM both viability and meiotic stages were evaluated. The results indicate no significant differences between procedures A and B with SOPS in all maturation stages, reaching a maximum maturation rate of 21%. As to Cryolock, significant differences were observed between both procedures, being procedure B, more efficient with a yield of 38% in MII stage and increased to 49% due to the co-culture with fresh granulosa cells. In conclusion, viability and maturation rates were improved with Cryolock and procedure B with the co-culture system in vitrified immature oocytes. (C) 2014 Elsevier Inc. All rights reserved.