两种分离斑马鱼卵母细胞方法的比较  

Comparison of Two Separating Methods of Zebrafish Oocytes Culture in Vitro

作　　者：吕众[1] 张毅[1] 魏华[1] 

机构地区：[1]上海海洋大学生命科学与技术学院,上海200090

出　　处：《生物技术通报》2008年第S1期342-346,355,共6页Biotechnology Bulletin

摘　　要：对培养斑马鱼卵母细胞体外成熟中的两种分离方法进行了比较。结果表明,在卵母细胞的分离过程中,机械分离法获得的完整卵细胞的比例(87.0%)显著多于EDTA-胰酶消化法(33.2%)(P<0.01)。但机械分离法较费时,其平均分离每个卵细胞所用的时间(0.69min)显著多于EDTA-胰酶消化法(0.17min)(P<0.01)。此外,经过18h 27℃培养,机械分离法获得的卵细胞的存活率(88.2%)显著高于EDTA-胰酶消化法所获卵细胞的存活率(60.1%),而采用机械分离法和消化分离法对所获卵细胞的成熟度-GVBD%分别为83.9%、77.5%,无显著差异(P>0.05)。基于数据所示,机械分离法可以获得存活率高、完整性好的卵细胞,但所需操作时间较长;EDTA-胰酶消化法操作便捷,但其分离出的卵细胞的活力较差;两种方法对卵细胞体外培养成熟率的影响无明显差异。The culture of zabrafish oocytes was studied by comparing the effects of two different separating methods—the mechanical method and EDTA-trypsin method—on the integrity,livability and GVBD%of the oocytes in vitro.After 18 hours incubation at 27℃the integrity and livability of the oocytes obtained by the mechanical method are 87.0%and 88. 2%.They are significantly higher than those obtained by EDTA-trypsin method(p<0.01),which are 33.2%and 60. 1%.But the mechanical method takes more time than EDTA-trypsin method(0.69 minute and 0.17 minute for an oocyte respectively).However,the GVBD%of the oocytes obtained by the two methods do not show obvious difference(p>0. 05)from the data(83.9%and 77.5%).The results show that the mechanical separating method had better effect on the integrity and livability of the oocytes than the EDTA-trypsin separating method,but it costs more work time.And the two methods didn't affect the GVBD%of the oocytes greatly,as the data shows.

关 键 词：斑马鱼 卵细胞 体外培养 机械分离 胰酶分离 比较 

分 类 号：Q813[生物学—生物工程]