GMP玻璃化冷冻未成熟牛卵母细胞核成熟及冷冻损伤试验  

Effect of GMP vitrification on bovine immature oocytes nuclear maturation and Cryodamage

作　　者：许卫华[1,2] 王学琼[1] 王子玉[1] 王锋[1] 张嘉保[3] 

机构地区：[1]南京农业大学动物胚胎工程技术中心,江苏南京210095 [2]福建省龙岩学院生命科学学院,福建龙岩364012 [3]吉林大学实验动物中心,吉林长春130062

出　　处：《中国兽医学报》2011年第6期917-920,932,共5页Chinese Journal of Veterinary Science

基　　金：国家科技支撑计划资助项目(2007BAD55B03)

摘　　要：本试验通过荧光染色的方法建立了未成熟牛卵母细胞在体外培养过程中第1次减数分裂的各个阶段的参考判定图谱;根据这个标准来观察毛细玻璃管(GMP)玻璃化冷冻对卵母细胞核成熟和冷冻损伤的影响。结果表明,从屠宰场废弃的卵巢表面卵泡内抽取的COCs,70%处于生发泡期,12.5%生发泡开始破裂,7.5%已开始浓缩,这说明从屠宰场获得的COCs有较高的异质性;卵母细胞在成熟培养22h时收获排出第一极体的卵母细胞,可得到丰度较高的极体-胞质染色体对称、紧密相邻的成熟卵母细胞;GMP玻璃化冷冻损伤主要有2种表现形式,首先,直接影响膜结构的完整性,包括细胞膜和核膜,这可从退化的细胞比例看出(8～24h,有21.9%～27.2%的细胞处于该阶段),其次,影响CONDENSED向MⅠ期的过渡,这可从处于CONDENSED卵母细胞的比例看出(8～24h,有24.1%～34.3%的细胞处于该阶段)。Firstly,fluorescent staining was used to establish the morphological standard of chromosomes,subsequently,based on the established standard;effect of freezing on nuclear maturation and cryodamage were analyzed.Results indicated：（1） at the beginning of fresh COCs in vitro maturation,70% COCs were found at GV atage,12.5% at GVBD stage,7.5% at CONDENSED stage,which demonstrated COCs from slaughterhouse were greatly heterogeneous;（2） after 8 hours of maturation,high-abundance of MⅡ oocytes having first polar body released and closely aligned in a line with cytoplasmic spindle;（3）GMP cryodamge presented in two types,among them,one presented as membrane damage including cell membrane and nuclear membrane,which could be demonstrated from 21.9%-27.2% degenerated oocytes during 8-24 h,the other presented as ceased CONDENSED→MⅠ transition,which could be demonstrated from 24.1%-34.3% oocytes at CONDENSED stage during 8-24 h.

关 键 词：牛 未成熟卵母细胞 毛细玻璃管 玻璃化冷冻 核成熟 损伤 

分 类 号：Q954.43[生物学—动物学]