目前建立的胚胎干细胞（embryonic stem cells, ESCs）被分为原始态（Naïve ）和 始发态（primed）两种状态，现在普遍认为来源大家畜的ES细胞大都处于primed状态。近年来的研究发现Wnt/β-Catenin信号通路是激活原始态ES细胞重要标志路径。本项目拟利用重编程方法结合胚胎干细胞技术探讨获取原始态多能性水牛ES细胞的方法；同时研究多能性水牛ES细胞中Wnt/β-catenin信号通路相关成分在mRNA和蛋白水平上的表达规律，并探讨Wnt/β-catenin通路激活剂（CHIR99021和BIO）及抑制剂（IWR-1-endo）对多能性水牛ES细胞培养效果和通路相关基因及蛋白表达的影响，从而阐明Wnt/β-catenin通路在ES细胞自我更新的调控作用。这将有助于丰富和完善水牛ES细胞自我更新的调控网络，对建立稳定的水牛ES细胞培养体系具有重要意义。

Embryonic stem cells（ESCs）currently are considering as two pluripotent states: naïve and primed. It is widely believed that most of embryonic stem cells (ESCs) from livestock are primed state. In recent years, the studies have indicated that Wnt/β-Catenin signal pathway plays an important role in activating the naïve embryonic stem cells. In this project, firstly, the possibility of obtaining the naïve buffalo ESCs through cells reprogramming and combining with the technology of embryonic stem cells are investigated. Secondly, the expression pattern of target genes mRNA and protein of the Wnt/β-Catenin signal pathway in the buffalo pluripotent ESCs are examined. Synchronously, the effects of activators (CHIR99021 and BIO) and inhibitor (IWR-1-endo) of Wnt/β-Catenin signaling pathway on the culture efficiency, as well as the expression of target genes and the proteins of Wnt/β-Catenin signaling pathway in buffalo pluripotent ESCs are also detected. This will help to clarify the function of Wnt/β-Catenin signaling pathway in the self-renewal of buffalo ES cells, enrich and consummate the regulation network of self-renewal in buffalo ES cells, so that to optimize the culture system of buffalo ES cells, and successfully establish bonafide ES cells line in buffalo.