氧分压、培养基及微孔对小鼠胚胎体外发育的影响  

Effects of Oxygen Tension,Medium and Well of the Well on In vitro Development of Mouse Embryo

作　　者：曹祖兵[1,2] 王维[1,2] 陆文昊[1,2] 周世贤[1,2] 李运生[1,2] 陶勇[1,2] 章孝荣[1,2] 张运海[1,2] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036 [2]安徽地方畜禽遗传资源保护与生物育种省级实验室,合肥230036

出　　处：《农业生物技术学报》2010年第5期956-962,共7页Journal of Agricultural Biotechnology

基　　金：国家科技支撑计划(No.2008BADB2B11-4);国家高技术研究发展计划(863)重点项目(No.2008AA101003)共同资助

摘　　要：通过小鼠(Mus musculus)胚胎体外培养,研究了培养基与不同氧分压环境之间是否存在偏好性;同时利用人体呼出的生理混合气体(肺气)对小鼠胚胎体外培养的可行性进行了探索;还比较了微孔(well of the well,WOW)培养法和成组培养法对胚胎发育的影响。结果表明:①在4%CO2+16%O2+78%N2+2%H2O(肺气)、5%CO2+5%O2+90%N(25%O2,低氧)和5%CO2+20%O2+75%N(220%O2,高氧)三种气相条件下,CZB和mKSOM两种培养基之间的胚胎卵裂率及囊胚率均差异不显著(P>0.05),但是,在肺气组中,CZB组的囊胚平均总细胞数明显高于mKSOM组(P<0.05);②低氧条件下,mKSOM组的囊胚率(22.6%)显著高于高氧和肺气条件下mKSOM组的囊胚率(P<0.05);③CZB培养基在低氧条件下获得的囊胚率与高氧或肺气条件下所获囊胚率差异不显著(P>0.05);④WOW培养法的囊胚发育率(74.6±5.1%)和囊胚平均总细胞数(76±2)显著高于成组培养法(分别为38.2±6.6%和58±4)(P<0.05)。结果提示:培养基mKSOM和CZB在支持胚胎发育时对氧分压具有偏好性;肺气能有效支持胚胎的正常发育;WOW培养法有利于提高胚胎体外发育能力和囊胚质量。The objectives of the present research are to investigate whether embryo culture media had preferences to oxygen tension during the in vitro development of mouse early embryos,and to explore feasibility of using physical lung air to support the in vitro development of mouse embryos,and to evaluate effect of well of the well（WOW） culture on in vitro pre-implantational deveopment of mouse embryos.The results were：First,cleavage rate and blastocyst rate were not significantly different between medium CZB and mKSOM on the use of three gas phases：4%CO2＋16%O2＋78%N2＋2%H2O（lung air）,5%CO2＋5%O2＋90%N2（5%O2,low oxygen） and 5%CO2＋20% O2＋75%N2（20%O2,high oxygen）（P0.05）,but mean total cell numbers per blastocyst cultured in CZB medium were higher than that in mKSOM when the lung air was used（P0.05）.Second,based on mKSOM medium,the blastocyst rate（22.6%） in 5%O2 gas phase was notably higher than that in other two gas phase（P0.05）.Third,as forCZB medium,blastocyst rate was not different significantly among three gas environments（P0.05）.Fourth,both the blastocyst rate（74.6±5.1%）and the mean total cell numbers per blastocyst（76±2）cultured in WOW system were obviously higher than that in the group culture system（38.2±6.6%and 58±4）.Taken together,these results indicate that the mKSOM medium and the CZB medium have their corresponding preferences to oxygen tension during in vitro culture of mouse embryos,and the lung air was reaffirmed to be able to effectively support in vitro pre-implantation development of mouse embryos,and WOW culture system can apparently enhance the in vitro developmental competence and blastocyst quality of mouse embryos.

关 键 词：小鼠胚胎 氧分压 培养基 偏好性 微孔培养法 

分 类 号：S823.36[农业科学—畜牧学]