本研究拟探讨原始态（naive）和始发态（primed）两种重编程状态水牛诱导多能干细胞（biPSC）的生物学特性、维持多能状态的机理、DNA甲基化修饰和嵌合能力等的差异，并将biPSC用于生产转基因克隆水牛。使用水牛转录因子Oct4、Sox2、Klf4、c-Myc结合水牛microRNA302/367重编程水牛体细胞为原始态和始发态的biPSCs。构建表达水牛bFGF和LIF的载体，使用水牛源bFGF和LIF诱导条件培养基对biPSCs培养建系。对两种状态水牛biPSC进行差异鉴定，主要包括培养条件、多能性、基因表达谱、维持各自多能性主要信号路径、X染色体失活、多能性基因启动子区甲基化水平、分化潜能和形成嵌合体能力等。探索两种状态的biPSC用于转基因克隆水牛生产的技术体系，鉴定iPSC克隆胚胎发育潜能。这些研究将为水牛干细胞的建系和应用于水牛遗传改良提供重要的理论和实践依据。

In this study, we focus on study the differences between na?ve and primed buffalo iPSC in biology characterization ,mechanism of maintaining the pluripotent state, DNA methylation modification and the ability to contribute to chimera, and using buffalo iPSC for generation trangenic-cloned buffalo. Buffalo transcription factors Oct4, Sox2, Klf4, c-Myc and miRNA302/367 will be used to generate the na?ve and primed buffalo iPSC. To establish the buffalo iPSC cell lines, the lentivirus vectors expressed LIF and bFGF will be constructed, and used for generation of LIF and bFGF-conditioned iPSC culture medium. The identification of na?ve and primed buffalo iPSC include the culture condition, the charateristics of pluriptency, gene expression profiling, the pluripotency-related core signaling pathways, X chromosome inactivation, the methylation status of the pluripotency -related genes, the differentiation potential in vitro and in vivo,and the ability to contribute to chimera. At last, how to use the two types of buffalo iPSC for generation transgenic-cloned buffalo will be investigated, and the developmental potential of iPSC derived-embryos will be identified.These efforts will provide important theoretical and practical basis for establishing buffalo pluripotent stem cell line and buffalo genetic improvement.