小鼠体外受精及其胚胎体外培养的比较研究  

The Comparative Studies on in vitro Fertilization and Development of Embryos in Mice

作　　者：贾青 高娟[1,2] 杨博[2,3] 刘丽均[2] 郁丽丽[2] 徐平[2] 芮荣[1] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]中国科学院上海实验动物中心,上海201615 [3]西北农林科技大学动物医学院,杨凌712100

出　　处：《实验动物与比较医学》2008年第5期304-308,共5页Laboratory Animal and Comparative Medicine

基　　金：国家十一五科技支撑项目(2006BAI23B04)

摘　　要：目的通过比较不同品系小鼠体外受精情况以及不同培养液对C57BL/6J小鼠胚胎体外培养效果的实验,进一步完善小鼠体外受精和早期胚胎体外培养体系。方法对C57BL/6J、DBA/2、FVB/NJ、BALB/c、KM、ICR及BARRⅡ小鼠进行超排和体外受精,并对C57BL/6J小鼠体外受精后的2-细胞胚胎用HTF、CZB、KSOM、M16、HECM五种培养液在相同条件下进行体外培养到囊胚,计算4-细胞～囊胚的发育率。结果各品系小鼠平均排卵数为13.0～38.7个(P<0.01),体外受精率为70.2%～92.8%(P<0.05)。不同培养液至4-细胞的发育率为60.3%～95.8%,其中KSOM的4-细胞发育率为95.8%,最适合2-细胞向4-细胞的发育。8-细胞发育率为52.8%～91.1%,桑椹胚发育率为40.6%～87.5%,囊胚发育率为5.1%～75.4%,各阶段胚胎发育率均差异显著。其中适合于金黄仓鼠桑椹胚、囊胚发育的HECM-2并不支持C57BL/6J囊胚的发育,其囊胚发育率只有5.1%,CZB最适合其囊胚的发育。结论遗传背景是影响小鼠超排及体外受精效果的主要因素之一。葡萄糖的存在不利于克服2-细胞发育阻断,胚胎在8-细胞期以前,主要能源物质不是葡萄糖,直到桑椹胚后期,葡萄糖才是主要的能量物质。磷酸盐可引起胚胎发育阻断。氨基酸和EDTA有利于克服胚胎发育阻滞。Objective To compare the fertilization in vitro in various mouse strains and the effects of different culture medium for development of embryos in C57BL/6J mice in vitro. Methods Females from 7 strains of mice were superovulated and operated by IVF, and the average number of superovulation and the fertility rate were observed. HTF, CZB, KSOM, M16, HECM were used as the culture medium to culture the2-cell of C57BL/6J mice in the same condition and the development rate from 4-cell to blastcyst were observed respectively. Results there is a significant difference （P〈0.01） in the average number of superovulation markedly among 13.0-38.7, and the rate of fertility of individual mouse with a range of 70.2% to92.8% （P〈0.05）. The development rate of 4-cell, 8-cell, morula and blastcyst in the 5 culture mediums with a range of 60.3%-95.8%, 52.8-91.1%, 40.6-87.5% and 5.1%-75.4%. Conclusion the genetic background is a primary factor for the effect of superovulation and fertilization in vitro in mice. Glucose can result in 2-cell interdiction of the embryo development; it is not primary energy substance before 8-cell. Phosphate can result in retardarce of the embryo development. Amino and EDTA are beneficial for overcoming the block of the embryo development.

关 键 词：小鼠 体外受精 胚胎体外发育 

分 类 号：Q813[生物学—生物工程]