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| Effect of cPouV and cNanog Down-regulation on Pluripotency of Chicken (Gallus gallus) Embryonic Stem Cells (cESCs) |
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Abstract To explore the pluripotent maintenance and update the functional influence of pluripotent genes cNanog and cPouV in chicken (Gallus gallus) embryonic stem cells(cESCs), cESCs were transfected by the stable RNAi vectors pSuper-cNanog and pSuper-cPouV constructed and screened early in our lab. Interference efficiency was detected with Real- time PCR and reached more than 65% at 96 h (P＜0.05). With downregulation of cNanog or cPouV gene expression, cESCs showed differentiation and growth rate of these cells slowed down. Domed colony of these cells gradually disappeared when the edge of colony became irregular. At 96 h after transfection, alkline phosphatase(AKP) and stage-specific embryonic antigen-1(SSEA-1) expression in cESCs were not be detected if cNanog gene was interfered but still stayed a part of expression in cells that cPouV gene was interfered. While downregulation of cNanog plus to cPouV gene expression, the remaining expressions of AKP and SSEA-1 were all disappeared. Results showed that cPouV and cNanog gene played an important role in maintaining pluripotency and self-renewal in chicken embryonic stem cells, and cNanog gene was dominant. These results may provide insights into the molecular regulation mechanism of avian during development and the difference between avian and mamal.
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Received: 10 March 2014
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