Four recombinant pluripotency transcriptional factors containing a protein transduction domain maintained the in vitro pluripotency of chicken embryonic stem cells

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• 作者：MiaoYing Yu (1) (2)
  Song Lian (1) (2)
  HongBing Han (2)
  Kun Yu (1) (2)
  GuiGuan Li (2)
  ZhengXing Lian (1) (2)
  Ning Li (1)
  
• 关键词：recombinant pluripotency factors ; protein transduction domain ; chicken ESC ; pluripotency
• 刊名：Science China Life Sciences
• 出版年：2013
• 出版时间：January 2013
• 年：2013
• 卷：56
• 期：1
• 页码：40-50
• 全文大小：1695KB
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• 作者单位：MiaoYing Yu (1) (2)
  Song Lian (1) (2)
  HongBing Han (2)
  Kun Yu (1) (2)
  GuiGuan Li (2)
  ZhengXing Lian (1) (2)
  Ning Li (1)
  
  1. National Key Laboratory of AgroBiotechnology, China Agricultural University, Beijing, 100193, China
  2. Laboratory for Molecular and Cellular Genetics, Collage of Animal Science and Technology, China Agriculture University, Beijing, 100193, China
  

文摘

Long-term in vitro maintenance of embryonic stem cell (ESC) pluripotency enables the pluripotency and differentiation of ESCs in animals to be investigated. The ability to successfully maintain and differentiate chicken embryonic stem cells (cESCs) would provide a useful tool for avian biology research and would be a resource directly applicable to agricultural production. In this study, endogenous chicken pluripotency transcription factors, POUV, Sox-2, Nanog and Lin28 were cloned and expressed as recombinant proteins containing a nine consecutive arginine protein transduction domain (PTD). cESCs were cultured with these recombinant proteins to maintain cESC pluripotency in vitro. Cultured cESCs exhibited typical characteristics of pluripotency, even after six generations of rapid doubling, including positive staining for stage-specific embryonic antigen I, and strong staining for alkaline phosphatase. Expression levels of the pluripotency markers, POUV, Nanog, C-Myc, Sox-2 and Lin28 were the same as in uncultured stage X blastoderm cells, and most significantly, the formation of embryoid bodies (EBs) by 6th generation cESCs confirmed the ability of these cultured cESCs to differentiate into cells of all three embryonic germ layers. Thus, transcription factors could be translocated through the cell membrane into the intracellular space of cESCs by using a PTD of nine consecutive arginines and the pluripotency of cESCs could be maintained in vitro for at least six generations.