畜牧兽医学报2017，Vol.48Issue(12)：2314-2322,9.DOI:10.11843/j.issn.0366-6964.2017.12.011
过表达Gata6诱导分离猪胚外内胚层干细胞样细胞
Porcine Extraembryonic Endoderm Stem Cell like Cells Induced by Overexpression of Gata6
摘要
Abstract
In this study,the role of Gata6 in porcine induced pluripotent stem cell (iPS cell) and extraembryonic endoderm stem cell (XEN cell) isolation was analyzed deeply.The required genes were cloned using templates of cDNAs,which were reverse transcripted from the mRNAs of the fetal tissues of Nongdaxiang pigs,and the retroviral plasmids pMXs-pOct4,pMXs-pSox2,pMXs-pKlf4,pMXs-pMyc,pMXs-pGata6,pMXs-pLin28,pMXs-pTbx3 and pMXs-pNanog were constructed,then,porcine fetal fibroblasts were infected with different combinations of genes.The results showed that co-transfection of Gata6 with Oct4,Sox2,Klf4 and mMyc (G6OSKM) could promote early reprogramming efficiency.Gata6 replacement of Oct4,with Sox2,Klf4 and mMyc (G6SKM),also could induce porcine fetal fibroblasts to iPS cells,but the reprogramming efficiency decreased,compared with the control group of OSKM (Oct4,Sox2,Klf4 and mMyc).Over expression of Gata6 forced pig iPS cells to undergo differentiation at the late stage of repro gramming,and changed into XEN cell like cells morphologically.These cells could maintain the morphology of XEN cells in mouse XEN cell culture medium,express XEN cell marker genes such as Gata4,Gata6 and Sox17.Overexpression of Gata6 in porcine iPS cells obtained by OSKM 4 factors also produced XEN cell like cells.The results indicate that overexpression of Gata6 can generate porcine XEN cell like cells,can provide theoretical basis for the isolation and derivation of porcine XEN cells from normal embryos and the exploration of the mechanisms of porcine early embryo developmental regulation.关键词
Gata6/诱导多能干细胞/胚外内胚层干细胞/重编程Key words
Gata6/induced pluripotent stem cells/extraembryonic endoderm stem cell (XEN cell)/reprogramming分类
农业科技引用本文复制引用
张伟,张少鹏,李恩宏,曹安,刘志宇,韩建永,曹素英..过表达Gata6诱导分离猪胚外内胚层干细胞样细胞[J].畜牧兽医学报,2017,48(12):2314-2322,9.基金项目
国家自然科学基金青年项目(31601941) （31601941）
北京市自然科学基金(6152004) （6152004）
北京市教委项目(PXM2016_014207_000008) （PXM2016_014207_000008）
北京市大学生科研训练项目(5046516564/003) （5046516564/003）
2015科研水平提高经费-高级别科研项目培育(1086716274) （1086716274）
兽医学(中医药)北京市重点实验室科研培育项目 （中医药）
