过表达Gata6诱导分离猪胚外内胚层干细胞样细胞

doi:10.11843/j.issn.0366-6964.2017.12.011

Abstract

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 reprogramming, 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.

CLC Number:

S828.2

ZHANG Wei, ZHANG Shao-peng, LI En-hong, CAO An, LIU Zhi-yu, HAN Jian-yong, CAO Su-ying. Porcine Extraembryonic Endoderm Stem Cell like Cells Induced by Overexpression of Gata6[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(12): 2314-2322.

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Porcine Extraembryonic Endoderm Stem Cell like Cells Induced by Overexpression of Gata6

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