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从山羊多能干细胞体外生成滋养层样干细胞。

In vitro generation of trophoblast like stem cells from goat pluripotent stem cells.

机构信息

The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, Inner Mongolia University, 010020, Hohhot, China; Research Center for Animal Genetic Resources of Mongolia Plateau, College of Life Sciences, Inner Mongolia University, 010020, Hohhot, China; College of Basic Medicine, Inner Mongolia Medical University, Hohhot, China.

The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, Inner Mongolia University, 010020, Hohhot, China; Research Center for Animal Genetic Resources of Mongolia Plateau, College of Life Sciences, Inner Mongolia University, 010020, Hohhot, China.

出版信息

Theriogenology. 2024 Sep 15;226:120-129. doi: 10.1016/j.theriogenology.2024.05.036. Epub 2024 May 24.

Abstract

Since the first mouse induced pluripotent stem cells (iPSCs) was derived, the in vitro culture of domestic iPSCs functionally and molecularly comparable with mouse iPSCs has been a challenge. Here, we established dairy goat iPSCs (giPSCs) from goat ear fibroblast cells with mouse iPSCs morphology, the expression of pluripotent markers and differentiation ability in vitro delivered by piggyBac transposon with nine Dox-inducible exogenous reprogramming factors. These reprogramming factors were bOMSK (bovine OCT4, CMYC, SOX2, and KLF4), pNhL (porcine NANOG and human LIN28), hRL (human RARG and LRH1), and SV40 Large T. Notably, AF-giPSCs (induced in activin A and bFGF condition) were capable of differentiation in embryoid bodies in vitro and could contribute to interspecies chimerism in mouse E6.5 embryos in vitro, demonstrating that AF-giPSCs have the developmental capability to generate some embryonic cell lineages. Moreover, Wnt/β-catenin signaling has an important role in driving goat induced trophoblast-like stem cells (giTLSCs) from Dox-independent giPSCs. This study will support further establishment of the stable giPSC lines without any integration of exogenous genes.

摘要

自从第一只小鼠诱导多能干细胞(iPSCs)被分离出来以来,体外培养与小鼠 iPSCs 在功能和分子水平上具有可比性的国产 iPSCs 一直是一个挑战。在这里,我们从山羊耳成纤维细胞中建立了奶山羊 iPSCs(giPSCs),这些细胞具有小鼠 iPSCs 的形态,通过带有 9 个 Dox 诱导的外源重编程因子的 piggyBac 转座子表达多能标记物和体外分化能力。这些重编程因子是 bOMSK(牛 OCT4、CMYC、SOX2 和 KLF4)、pNhL(猪 NANOG 和人 LIN28)、hRL(人 RARG 和 LRH1)和 SV40 大 T。值得注意的是,AF-giPSCs(在激活素 A 和 bFGF 条件下诱导)能够在体外胚胎体中分化,并能够在体外的小鼠 E6.5 胚胎中产生种间嵌合体,表明 AF-giPSCs 具有产生一些胚胎细胞谱系的发育能力。此外,Wnt/β-catenin 信号通路在驱动 Dox 非依赖性 giPSCs 产生诱导滋养层样干细胞(giTLSCs)方面起着重要作用。本研究将支持进一步建立稳定的 giPSC 系,而无需任何外源基因的整合。

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