摘要
对水牛和小鼠腔前卵泡的分离方法进行了系统探索,旨在找出一种水牛和小鼠腔前卵泡的有效分离方法。实验一分别用梳刮法、剪碎法和显微分离法分离回收水牛卵巢腔前卵泡;实验二采用梳刮法分离水牛胎牛、青年牛和成年牛腔前卵泡;实验三分别用浓度为0(CK)、0.02%、0.04%、0.08%的胶原酶消化水牛卵巢20 m in,然后采集腔前卵泡;实验四采用梳刮法、剪碎法和显微分离法分离小鼠卵巢的腔前卵泡。结果表明,梳刮法能有效地分离回收水牛和小鼠卵巢的腔前卵泡。该法分离水牛卵巢所得腔前卵泡的原始卵泡最多,次级卵泡最少,而分离小鼠卵巢所得腔前卵泡的次级卵泡最多,原始卵泡最少;水牛胎牛卵巢可分离得到较多的腔前卵泡;酶消化有助于提高剪碎法的腔前卵泡分离效率。
A tool named comb-scrape which consists of needles, a needle fixing and a handle was designed to investigale the methods for isolation of buffalo and mouse preantral follicles were investigated in this study.
Comb-scrape method harvested significantly more buffalo preantral follicles [ (152.35±44.81) / ovary] than the methods of cutting [ (32.62± 14.81) /ovary] and micromanipulation [ (8.95 ± 3.44) /ovary, P〈0.05]. The average manipulative time for each ovary (39.05±4.27) min was also less than that of the methods of cutting (46.43±4.15) min and micromanipulation [(44.55±7.82) min, P〈0.05]. Majority of preantral follicles isolated by comb-scrape method were primordial follicles (41.25% ) and primary follicles (38.79 % ), and very few of secondary follicles (19.95 %) . These results indicated that comb-scrape method was suitable for isolating buffalo preantral follicles.
Fetal buffalo ovaries harvested significantly more preantral follicles [ (943.43±143.13) /ovary] than heifer [ (249.50±40.11) /ovary] and adult ovaries [ (128.50±19.99) /ovary] (P〈0.01), indicating that more preantral follicles can be isolated from buffalo fetal ovaries. Digestion of small buffalo ovary tissue pieces with 0.02%, 0.04% and 0.08% collagenase harvested significantly more preantral follicles in comparison with without digestion (40.38 ±6.12, 47.75±6.84and 52.69±6.15 vs 33.56±6.15 per ovary, P〈0.05), indicating that combination of cutting and enzymatic method can harvest more buffalo preantral follicles than cutting method alone.
Comb-scrape method harvested significantly more mouse preantral follicles [(61.00 ± 9.12)/ ovary] than the methods of cutting [(26.30±5.06)/ovary] and micromanipulation [(14.10±4.16)/ ovary,P〈0.05].The average manipulative time for each ovary (30.80±3.91)rain was also less than that of the methods of cutting (39.10±4.01)min and micromanipulation [(38.40±3.50)min,P〈 0.05]. Majority of preantral follicles isolated by comb-scrape method were secondary follicles (83.61%) and primary follicles (12.13%),and very few of primordial follicles (4.26%). These results indicate that comb-scrape method is also suitable for isolating mouse preantral follicles.
出处
《广西农业生物科学》
CAS
CSCD
2006年第B09期197-197,199,共2页
Journal of Guangxi Agricultural and Biological Science
基金
国家自然科学基金项目(30460090)
广西科学基金项目(桂科自0640016)
广西大学博士启动项目(X041110)
关键词
腔前卵泡
分离
水牛
小鼠
preantral follicle
isolation
buffalo
mouse