SC1(pluripotin)对昆明小鼠胚胎干细胞分离培养的影响  

Effects of SC1(pluripotin) on isolation and culture of embryonic stem cells from Kunming mouse

作　　者：任卫青[1] 张志平[1] 王新庄[1] 吕婧玉[1] 许晓婷[1] 蒋金航[1] 翟明胜[1] 石凯文[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450001

出　　处：《中国兽医学报》2013年第7期1132-1136,共5页Chinese Journal of Veterinary Science

基　　金：河南省基础与前沿技术研究计划资助项目(092300410081);河南农业大学人才引进基金资助项目

摘　　要：比较了在无血清培养体系中分别添加小分子化学物质pluripotin(SC1)和白血病抑制因子(LIF)对昆明小鼠ES细胞未分化状态的维持作用,并探讨了SC1在培养液中的最佳浓度。以小鼠胎儿成纤维细胞(MEF)为饲养层,以含LIF的添加血清替代品(KSR)的培养基(ESC-SR)为对照,分别用SC1浓度为0.3、1、3μmol/L的ESC-SR培养液对昆明小鼠胚胎进行分离培养。结果显示:胚胎在添加3μmol/L SC1的ESC-SR培养液中ICM形成率显著高于对照组(P<0.05),形成的ES集落与对照组形态差异不明显,最高传至第7代,并且在无饲养层条件下可以传至第6代而保持未分化状态。不同浓度SC1培养液中,胚胎在SC1浓度为0.3μmol/L组的F2代形成率显著高于1μmol/L组和3μmol/L组(P<0.05),所分离的细胞AKP染色呈阳性,Oct4、Nanog和Sox2的免疫荧光染色均显阳性,具有ES细胞的特点。结果表明,ESC-SR培养液中SC1可以替代LIF用于小鼠ES细胞未分化状态的维持,并且可以在无饲养层条件下进行昆明小鼠ES细胞的培养。In order to study the effection of the small molecules（SC1） in maintaining embryonic stem cell self-renewal state,the experiment investigated formation of ICM outgrowths and isola- tion of embryonic stem cells from mouse embryos of Kunming specie, which were used as the starting material while mitomycin C-inactivated MEF as the feeder layer. In the serum-free medi- um added with SC1 in different concentration of 0.3,1 and 3 μmol/L instead of LIF and LIF add- ed as control. The results demonstrated that the formation rates of ICM outgrowths were signifi- cant highter than control group,and the second passage rate in the medium added with 0.3 μmol/ L SC1 were higher than added with 1 μmol/L and 3 μmol/L,the ES cells maintained undifferentiat- ing for 7 passages, and maintained undifferentiating for 6 passages without feeder layer cells. The isolated cells were positive for AKP staining and immunofluorescence analysis positive against antigens of Oct4, Nanog and Sox2. In conclusion, SC1 can maintain the pluripotency and undifferentiation of ES cells from Kunming mouse without LIF and feeders, cells.

关 键 词：昆明小鼠 SC1 胚胎干细胞 分离培养 

分 类 号：S814.8[农业科学—畜牧学]