活体采卵时间间隔对牛克隆胚胎发育的影响  

Intervals of OPU Affect the Quality of Bovine Oocytes Used for SCNT

作　　者：王敬军[1] 田海滨 丁利军[3] 陈娟[1] 徐旭俊[1] 沙红英[1] 陈建泉[1] 成国祥[1] 

机构地区：[1]同济大学生命科学与技术学院,上海200092 [2]上海转基因研究中心,上海201203 [3]上海交通大学医学院附属瑞金医院,上海200025

出　　处：《上海交通大学学报（农业科学版）》2008年第1期46-49,77,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

摘　　要：采用活体穿刺技术(ovum pick up,OPU),分别间隔3/4(1周2次)、5、7、10、14 d对同一批荷斯坦奶牛进行穿刺采卵,根据优势卵泡直径和所有卵丘细胞-卵母细胞复合体的状态确定采卵时卵泡所处的发育阶段,从而确定卵泡波的发育动态变化。并以间隔不同时间采卵获得的卵母细胞成熟后进行体细胞核移植(somatic cell nuclear transfer,SCNT)制备克隆牛胚胎,重构胚体内培养7 d后,通过比较克隆囊胚的发育率判定卵母细胞质量的差异,从而确定最佳的采卵时间间隔。结果显示:①穿刺采集所有直径超过3mm的卵泡后,可起始1个新卵泡波,大约5 d后从属卵泡开始进入凋亡阶段,7 d后进入晚期凋亡阶段,第7～10 d又起始1个新的卵泡波,第14 d时第2个卵泡波中的从属卵泡起始凋亡。②间隔不同时间组克隆囊胚发育率存在差异,间隔3/4 d(1周2次)为23.1%,间隔5 d组为15.0%,间隔7 d组为10.9%,间隔10 d组为4.9%,间隔14 d组为29.0%。结果显示:间隔不同的时间采卵,卵泡所处的发育状态存在差异,并对卵母细胞质量具有显著的影响。Cow follicular oocytes were aspirated from the ovaries of Holsteins by ovum pick up （OPU） at intervals of 3/4（twice/week）, 5, 7, 10, 14 days, respectively, The developmental stages of follicles were confirmed by state of dominant follicles and cumulus-oocytes complexes, subsequently, the dynamics of follicular wave was clarified. In addition, after IVM, oocytes were used to produce cloned bovine embryos which were then cultured in vivo for 7 days, and the developmental rates of blastocysts were compared between the five experimental groups. The proper OPU interval was confirmed by the developmental rates of cloned blastocysts. The results showed that a new follicular wave starts after all follicles whose diameter were beyond 3 mm were picked up, subordinate follicles will become atretic 5 days later, another follicular wave starts between day 7 and 10 and the subordinate follicles in second follicular wave become early atretic at day 14. The developmental rates of cloned blastocysts were 23.1% （3 or 4 day interval, twice/week）, 15.0% （5 day interval）, 10.9% （7 day interval）, 4.9% （10 day interval）, 29% （14 day interval）, respectively. The results indicate that the developmental stages of follicles are different when different OPU intervals were adopted, which affects the quality of oocytes.

关 键 词：奶牛 活体采卵 体细胞核移植 卵母细胞质量 

分 类 号：Q954.4[生物学—动物学]