A novel strategy to derive iPS cells from porcine fibroblasts  

作　　者：RUAN WeiMin HAN JianYong LI Pin CAO SuYing AN Yang LIM Bing LI Ning 

机构地区：[1]State Key Laboratory for Agrobiotechnology,College of Biological Sciences,China Agricultural University,Beijing 100193,China [2]Stem Cell and Developmental Biology,Genome Institute of Singapore,138672,Singapore

出　　处：《Science China(Life Sciences)》2011年第6期553-559,共7页中国科学（生命科学英文版）

基　　金：supported by the National Basic Research Program of China (Grant Nos. 2009CB941003, 2011CBA0110 and 2011CBA01000)

摘　　要：Induced pluripotent stem (iPS) cell technology demonstrates that somatic cells can be reprogrammed to a pluripotent state by over-expressing four reprogramming factors.This technology has created an interest in deriving iPS cells from domesticated animals such as pigs,sheep and cattle.Moloney murine leukemia retrovirus vectors have been widely used to generate and study mouse iPS cells.However,this retrovirus system infects only mouse and rat cells,which limits its use in establishing iPS cells from other mammals.In our study,we demonstrate a novel retrovirus strategy to efficiently generate porcine iPS cells from embryonic fibroblasts.We transfected four human reprogramming factors (Oct4,Sox2,Klf4 and Myc) into fibroblasts in one step by using a VSV-G envelope-coated pantropic retrovirus that was easily packaged by GP2-293 cells.We established six embryonic stem (ES)-like cell lines in human ES cell medium supplemented with bFGF.Colonies showed a similar morphology to human ES cells with a high nuclei-cytoplasm ratio and phase-bright flat colonies.Porcine iPS cells could form embryoid bodies in vitro and differentiate into the three germ layers in vivo by forming teratomas in immunodeficient mice.Induced pluripotent stem （iPS） cell technology demonstrates that somatic cells can be reprogrammed to a pluripotent state by over-expressing four reprogramming factors.This technology has created an interest in deriving iPS cells from domesticated animals such as pigs,sheep and cattle.Moloney murine leukemia retrovirus vectors have been widely used to generate and study mouse iPS cells.However,this retrovirus system infects only mouse and rat cells,which limits its use in establishing iPS cells from other mammals.In our study,we demonstrate a novel retrovirus strategy to efficiently generate porcine iPS cells from embryonic fibroblasts.We transfected four human reprogramming factors （Oct4,Sox2,Klf4 and Myc） into fibroblasts in one step by using a VSV-G envelope-coated pantropic retrovirus that was easily packaged by GP2-293 cells.We established six embryonic stem （ES）-like cell lines in human ES cell medium supplemented with bFGF.Colonies showed a similar morphology to human ES cells with a high nuclei-cytoplasm ratio and phase-bright flat colonies.Porcine iPS cells could form embryoid bodies in vitro and differentiate into the three germ layers in vivo by forming teratomas in immunodeficient mice.

关 键 词：induced pluripotent stem cells Moloney murine leukemia retrovirus vectors embryoid body TERATOMA 

分 类 号：Q786[生物学—分子生物学] S865.13[农业科学—野生动物驯养]