昆明种小鼠胚胎干细胞的培养及鉴定.doc

昆明种小鼠胚胎干细胞的培养及鉴定.doc昆明种小鼠胚胎干细胞的培养及鉴定作者:高巍单位:辽宁医学院解剖学教研室,辽宁锦州121001【摘要】目的从昆明种小鼠的早期胚胎中获取并培养胚胎干细胞。,用小鼠胚胎成纤维细胞作为饲养层,形成的ES细胞样集落,72h后分离隆起生长的内细胞团并继续培养,观察集落的生长状态,并通过碱性磷酸酶染色进行鉴定。结果ES细胞有其典型的形态学特征:集落呈鸟巢状,边缘清楚,表面平滑,结构致密,隆起生长,细胞之间界限不清楚;单个细胞体积小、核大;对ES细胞碱性磷酸酶进行检测,在AKP底物作用下,未分化的ES细胞显微镜下为棕褐色,分化的不着色。结论昆明种小鼠囊胚在小鼠胚胎饲养层细胞上可以发育为胚胎干细胞。【关键词】小鼠胚胎干细胞ES细胞系Isolation,CultureandIdentificationofMurineEmbryonicStemCellsofKunmingSpeciesGAOWeil,LIUFuqiangl,MEIXifanl,LIUChang2,RENFu3(;,LiaoningMedicalUniversity;,LiaoningMedicalUniversity,Jinzhou121001China)Abstract:ObjectiveToisolateandcultureembryonicstem(ES),(ICM)-likecolonies,roundorelliptic,withclearedgeandsmoothsurface,strongrefraction,pactnessofcellaggregation,andobscuredistinctionbetweentwoedgesofneighbourcellswithinacolony,,alloftheEScellsshowedstrongAKPactivity,andonthecontrary,:Kunmingspec