摘要
本研究旨在探索建立猪GV（Germinal visiele）期卵母细胞的冷冻保存方法。用抽吸法采集屠宰猪卵巢中直径为2—5mm及〉5mm的卵泡卵母细胞，再用切割法采集直径〈2mm的卵泡卵母细胞。结果表明，切割法所获卵母细胞数及每只卵巢平均采卵数均显著高于抽吸法（P〈0．05）；但切割法所获卵母细胞可用率仅为33．8％，显著低于抽吸法67．5％的可用率（P〈0．05）。用抽吸法采集直径为2—5mm的卵泡卵母细胞，进行体外成熟和玻璃化冷冻研究。4组成熟培养结果表明，卵母细胞在添加激素和猪卵泡液（pFF）的成熟培养液中培养24h后转入无激素、无卵泡液的基础培养液中继续培养20h，体外成熟率达78．9％，显著高于另外3组（P〈0．05）。以EFS40作为玻璃化冷冻液，比较细管法和OPS（Open pulled straw）法对猪GV期卵母细胞的冷冻效果，从冻后形态正常率来看，两种方法间无统计学差异（P〉0．05）；但OPS法冷冻猪GV期卵母细胞的冻后存活率和体外成熟率均显著高于细管法（P〈0．05）。
The purpose of this experiment is to investigate and establish a vitrification method for porcine GV （ germinal visicle） stage oocytes. The aspiration method was used to collect oocytes from follicles 2--5 mm and 〉5 mm in diameter and the cutting method was adopted to collect oocytes of follicles 〈 2 mm in diameter from porcine ovaries obtained in slaughterhouse. The result showed that either total number or average number per ovary of oocytes obtained with the cutting method were significantly higher than those from the aspiration method （ P 〈0. 05）. However, the percentage of available oocytes from the cutting method was 33.8% and significantly lower than the percentage of the aspiration method （67. 5% ）. Oocytes of follicles 2--5 mm in diameter were aspirated for in vitro maturation and vitrification experiments. Oocytes were cultured in the maturation medium containing hormones and porcine follicular fluid （pFF） for 24 h and then transferred into the basic medium without hormones and pFF for further 20 h, resulting in 78. 9% of maturation rate, which was significantly higher than those from other 3 groups （ P 〈 0. 05 ）. The EFS40 was used as freezing solution and porcine GV stage oocytes were loaded in open pulled straw （OPS） and 0. 25 ml straw respectively for cryopreservation. These two vitrification methods yielded similar post-thawing normal rates in morphology （ P 〉 0. 05） but significantly different post-thawing survival rates and maturation rates in vitro （ P 〈 0. 05 ）, indicating that the OPS method was significantly superior to the straw method in post-thawing survival rate and maturation rate in vitro.
出处
《江苏农业学报》
CSCD
北大核心
2006年第2期131-136,共6页
Jiangsu Journal of Agricultural Sciences
基金
上海市科技兴农重点攻关项目(沪农科攻字2003第14-1号)
国家自然科学基金项目(30270958)
作者简介
武彩红（1976-），女，山西和顺人，博士研究生，研究方向为动物生殖生物学。
通讯作者：芮荣，（E-mail）rrui@njau．edu．cn；（Tel）025-84395595
