wnt3a蛋白对小鼠胚胎干细胞多能性维持以及诱导多能干细胞重编程效率的影响动物学专业论文.docxVIP

Wnt3a蛋白对小鼠胚胎干细胞多能性的维持以及诱导多能干细胞重编程效率的影响当在无饲养层条件培养ESCs时，L．Wnt3a细胞条件培养液可显著维持ESCs Wnt3a蛋白对小鼠胚胎干细胞多能性的维持以及诱导多能干细胞重编程效率的影响 当在无饲养层条件培养ESCs时，L．Wnt3a细胞条件培养液可显著维持ESCs 的克隆形态，而传统干细胞培养液中培养的ESCs克隆发生明显分化。L—Wnt3a 细胞条件培养液培养的ESCs的多能性基因sox2、oct4、nanog以及表观修饰基 因parpl、kdm2b的转录水平都显著高于对照组ESCs培养液培养的ESCs。而这 两组细胞的Wnt信号通路基因的表达则没有出现显著改变，相比分化较严重的 MEF培养液培养的ESCs，两组细胞cd31和birc5的转录水平相对较高。此外， L．Wnt3a细胞条件培养液培养的ESCs能够较好的维持干细胞的多能性，体外分 化形成的类胚体表达更高水平的三胚层标记基因，并体内分化形成畸胎瘤。 本研究显示，Wnt3a信号通路会显著促进ESCs多能性的维持并上调与ESCs 多能性维持相关的表观遗传修饰基因的表达。进而，该信号通路显著促进小鼠 iPSCs的诱导，并显著上调与多能性调控相关的表观遗传修饰基因的表达。本实 验证实，Wnt3a信号通路不但对多能性基因转录有显著的调控作用，而且也影响 与多能性相关的表观遗传调控基因的表达。该研究的结论为进一步阐明Wnt3a 信号通路在调控多能性，特别是诱导过程中多能性的获得奠定理论基础。 关键词：Wnt3a；无饲养层培养；多能性维持 万方数据 内蒙古大学 内蒙古大学 硕士学位论文 RESEARCH oF THE WNT3A FOR PLURIPoTENCY MAINTANCE oF STEM CELLS AND PRoMoTING THE EFFICIENCY oF REPRoGRAMMING ABSTRACT Wnt signaling pathway plays significant roles in embryonic differentiation and maintaining pluripotency of stem cells and SO on．This signaling pathway initiating complicated series of intracellular signal mediation by extracellular proteins and cell surface receptor binding，consequently regulating the expressions of relative genes． HoweveL the epigenefic modifications in maintaining and inducing the pluripotency of stem cells of Wnt signaling have not known yet． This study is aimed at using mouse embryonic stem cells(ESCs)and induced pluripotent stem cells (iPSCs) as expefimenml objectives， utilizing the function of secreting Wnt3a proteins to extracellular space of L—Wnt3a cells， preparing L--Wnt3a conditioned medium or take L·-Wnt3a cells as feeder layers to culture ESCs or induce iPSCs，in order to discuss the role of pluripotency maintenance of ESCs and inducing reprogramming iPSCs of Wnt signaling pathway． The results show that take L—Wnt3a cells as feeder layers of ESCs can maintain their pluripotent state well，this ESCs express alkaline phosphatase and other pluripotent 万方数据 Wnt3a蛋白对小鼠胚胎干细胞多能性的维持以及诱导多能干细胞重编程效率的影响relative Wnt3a蛋白对小鼠胚胎干细胞多能性的维持以及诱导多