小鼠原核期受精卵体外培养系统的筛选  

Filtration of Culture System in Vitro of Mouse Pronuclear Zygote

作　　者：淡新刚[1] 董雅娟[2] 柏学进[2] 马保华[1] 胡亮[1] 杨恕玲[2] 王雪红[2] 董方圆[2] 

机构地区：[1]西北农林科技大学家畜生殖内分泌与胚胎工程重点开放实验室,杨凌712100 [2]莱阳农学院动物胚胎工程中心,莱阳265200

出　　处：《生物技术通报》2006年第C00期416-421,共6页Biotechnology Bulletin

基　　金：青岛市科技将才资助项目

摘　　要：为了筛选出最佳的小鼠原核期受精卵的体外发育培养系统,分别进行了四个试验。试验I:在体外分别用自配的M16、mM16、KSOM、mKSOM、CZB进行体外发育培养,进而筛选出一种最佳的体外培养系统;试验Ⅱ、Ⅲ、Ⅳ分别:探讨了血清、PVA、rhLIF对小鼠胚胎发育的影响。结果,试验I中胚胎发育到2-细胞的比率差异不明显,但是在mM16和mKSOM中,发育到4-细胞的比率94.7%,90.7%(91/96;78/86)和发育到桑椹胚/囊胚的比率分别为78.1%,67.4%(75/96;58/86)均明显高于其他三种培养液;试验II用10%FBS代替M16中BSA时,胚胎的发育率均下降,即使在mM16中桑椹胚/囊胚率仅为4.8%(12/35)与对照组M16(40.5%)差异显著(p<0.05);试验Ⅲ用PVA取代mM16和mKSOM中的BSA其体外发育率显著下降,胚胎均无一例发育到桑椹胚/囊胚;试验Ⅳ:rhLIF能提高胚胎在体外的发育率可使mM16培养的胚胎囊胚率、囊胚脱出率分别达到84%(47/56)、39.2%(22/56)。结论:在不添减其他成分前提下,只在M16中添加0.1mMolEDTA、0.5mMol牛磺酸、1000IU/mlrhLIF便可获得84%的囊胚率,同时证明在M16或mM16添加血清都会降低其体外发育率;PVA还不能有效的取代mM16、mKSOM中的血清。To filtrate the best culture system in vitro of mouse pronuclear zygote, we carry through separately four experimentation. Text Ⅰ ,using respectively M16,mM16,KSOM, mKSOM,CZB which were confected by us carry through developmental culture in vitro on mouse pronuclear zygote and filtrate a best cultural system ;test Ⅱ, we discuss separately FBS,PVA,rLIF how to effect the development of mouse embryo. Result： test Ⅰ ,the difference of the ratio zygote develop 2- cell is unconspicuous, but the ratio zygote develop 4-cell,morula,blastocyst are abviously higher than else media; test Ⅱ , the viability of zygote all drops using 10% FBS replace BSA in M16 ,even in mM16 ,the ratio of morula or blastocyst is just 4.8% ,compared comparision group M16（40.5% ）, the different is distinct： test Ⅲ, the viability decline through PVA substitute BSA of mM16,mKSOM , none of embryos develop morula or blastocyst; test Ⅳ ,rhLIF can improve remarkablely the viability of embryo in vitro and the ratio of blastocyst and prolaspsing blastocyst from the zona pellucida can achieve separately 84% ,39.2% in mM16. Conclusion： under not appending or subtracting other components, appending merely 0.1 mMol EDTA,0.5 mMol taurine,1 000IU/ml rhLIF in M16, then make the ratio of blastacyst reach to 84%. Simultaneously proving： in M16 or mM16 appending FBS may reduce the viability of zygote in vitro, PVA can＇ t substitute effectively FBS in mM16,mKSOM.

关 键 词：小鼠 原核期受精卵 体外培养 筛选 

分 类 号：Q813[生物学—生物工程]