添加不同因子对绵羊类胚胎干细胞多能性的影响  

Effects of Different Regulators on oESC Pluripotency

作　　者：张秀华[1,2] 石庆华[1] 赵云程[2] 林嘉鹏[2] 汪立芹[2] 周川[1,2] 金贤华[1,2] 黄俊成[2] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]新疆畜牧科学院,新疆动物生物技术重点开放实验室,农业部草食家畜繁育生物技术重点开放实验室,乌鲁木齐830000

出　　处：《中国草食动物》2012年第3期25-29,共5页China Herbivores

基　　金：新疆维吾尔自治区高技术研究与发展计划(200711104);新疆维吾尔自治区高技术研究与发展计划(201111113);国家转基因重大专项(2011ZX08008-003)

摘　　要：为筛选出更有利于维持绵羊类胚胎干细胞(oESC-like cell)多能性的因子,在基础培养基N2B27/CH中,分别添加PD、PD+hLIF、PD+BPM4、PD+hLIF+BMP4,通过比较分析,初步筛选出了比本实验室原培养体系(N2B27/CH+bFGF)更利于维持oESC-like多能性的培养基添加因子。结果显示:基础培养基中添加PD、PD+hLIF后细胞生长较好,细胞之间的结合更加致密;各种培养基培养物都表达AKP与Sox2、Oct4免疫荧光蛋白等oESC-like多能性标志;4种不同培养基与bFGF相比,细胞的生长速度减慢;在基础培养基N2B27/CH中添加PD和PD+hLIF使多能性候选基因Lin28与c-Myc表达量极显著升高(P<0.01),Nestin的表达量极显著下调(P<0.01),加入PD+hLIF使Oct4表达量极显著上调(P<0.01)。因此,在oESC-like基础培养液N2B27/CH的基础上,添加PD或PD+hLIF更有利于oESC-like细胞多能性维持。In order to find the more effective regulators for maintaining the pluripotency of ovine embryonic stem-like cell （oESC）, the bFGF （control）, PD, PD ＋hLIF, PD ＋ BMP4 and PD ＋ hLIF ＋ BMP4 were added to the basal medium （N2B27） respectively. The results showed that the cells in the PD, PD＋hLIF groups grew better and more dense;The expression of AKP, Sox2 and Oct4 etc were observed in all groups,which were the pluripotency symbols of oESC;Compared with the bFGF group , the cell grew slowly in the other four groups;In PD and PD＋hLIF adding groups,the expression of Lin28 and c-Myc genes increased very significantly（P〈0.01 ）, while the expression of Nestin gene decreased very significantly （P〈0.01）;The expression of Oct4 gene increased extreme significantly （P〈0.01） in PD＋hLIf group.It was concluded that the addition of PD or PD＋hLiF to the basal medium would be more beneficial to the pluripotency maintainance of oESC.

关 键 词：绵羊类胚胎干细胞 体外培养 多能性调控因子 候选基因 

分 类 号：S826.3[农业科学—畜牧学]