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玻璃化对山羊卵丘-卵母细胞复合体减数分裂成熟和基因表达的影响。

Effect of vitrification on meiotic maturation and expression of genes in immature goat cumulus oocyte complexes.

机构信息

Laboratory for Conservation of the Endangered Species (LaCONES), Centre for Cellular and Molecular Biology (CCMB), Hyderabad, India.

出版信息

Cryobiology. 2012 Jun;64(3):176-84. doi: 10.1016/j.cryobiol.2012.01.005. Epub 2012 Jan 18.

Abstract

The aim of the study was to evaluate meiotic maturation, and expression of genes coding for oocyte secreted factors (GDF9, BMP15, TGFBR1, and BPR2) and apoptosis (BCL2, BAX and P53) after vitrification of immature goat cumulus oocyte complexes (COCs) and in vitro maturation. COCs were vitrified in a solution containing ethylene glycol, dimethyl sulfoxide and sucrose using either a conventional straw (CS), open pulled straw (OPS), cryoloop (CL), hemistraw (HS) or cryotop (CT). Freshly collected COCs (Control), COCs exposed to vitrification and dilution solutions without cryopreservation (EC) and vitrified-warmed COCs were matured in vitro for 27h. The viability of vitrified-warmed COCs 2 h post warming and in vitro maturation was similar for CL, HS and CT. The proportion of oocytes that extruded a 1st polar body and reached TI/MII was significantly higher with CT and HS followed by CL, OPS and CS. Gene expression of GDF9, BMP15, BMPR2, BAX and P53 were comparable to control levels for OPS, CL, HS and CT. The gene expression pattern in CS vitrified COCs was by contrast changed in that GDF9, BMP15, TGFBR1 and BAX were up regulated and BMPR2, BCL2 and P53 down regulated. In conclusion immature goat COCs vitrified using CT and HS showed that viability, maturation rates and expression of genes coding for oocyte secreted factors and apoptosis were similar to non-vitrified controls.

摘要

本研究旨在评估玻璃化冷冻对不成熟山羊卵丘-卵母细胞复合物(COCs)的减数分裂成熟和编码卵母细胞分泌因子(GDF9、BMP15、TGFBR1 和 BPR2)及凋亡(BCL2、BAX 和 P53)基因表达的影响,以及体外成熟的情况。采用含有乙二醇、二甲基亚砜和蔗糖的溶液,通过传统 straw(CS)、open pulled straw(OPS)、cryoloop(CL)、hemistraw(HS)或 cryotop(CT)对 COCs 进行玻璃化冷冻。新鲜采集的 COCs(对照)、未进行冷冻保存的暴露于玻璃化和稀释溶液中的 COCs(EC)和玻璃化冷冻后解冻的 COCs 进行体外成熟 27 小时。解冻后 2 小时和体外成熟后,CL、HS 和 CT 玻璃化冷冻解冻 COCs 的活力相似。CT 和 HS 后紧接着是 CL、OPS 和 CS,排出第一极体并达到 TI/MII 的卵母细胞比例显著更高。OPS、CL、HS 和 CT 的 GDF9、BMP15、BMPR2、BAX 和 P53 基因表达与对照水平相当。相比之下,CS 玻璃化冷冻 COCs 的基因表达模式发生了变化,GDF9、BMP15、TGFBR1 和 BAX 上调,BMPR2、BCL2 和 P53 下调。总之,使用 CT 和 HS 对不成熟山羊 COCs 进行玻璃化冷冻,其活力、成熟率和编码卵母细胞分泌因子及凋亡的基因表达与非玻璃化对照相似。

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