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成年成纤维细胞和羊水来源干细胞构建的水牛手工克隆胚胎植入前阶段发育重要基因表达谱的比较研究

A comparative study on expression profile of developmentally important genes during pre-implantation stages in buffalo hand-made cloned embryos derived from adult fibroblasts and amniotic fluid derived stem cells.

作者信息

Em Sadeesh, Shah Fozia, Kataria Meena, Yadav P S

机构信息

Division of Animal Physiology and Reproduction, ICAR-Central Institute for Research on Buffaloes, Hisar, 125001, India.

Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, 125004, India.

出版信息

Cytotechnology. 2016 Aug;68(4):1447-61. doi: 10.1007/s10616-015-9904-7. Epub 2015 Jul 30.

Abstract

Abnormal gene expression in somatic cell nuclear transfer embryos due to aberrant epigenetic modifications of the donor nucleus may account for much of the observed diminished viability and developmental abnormalities. The present study compared the developmentally important gene expression pattern at 4-cell, 8- to 16-cell, morula, and blastocyst stages of buffalo nuclear transfer (NT) embryos from adult fibroblasts (AFs) and amniotic fluid stem cells (AFSCs). In vitro fertilized embryos were used as control embryos. Alterations in the expression pattern of genes implicated in transcription and pluripotency (OCT4, STAT3, NANOG), DNA methylation (DNMT1, DNMT3A), histone deacetylation (HDAC2), growth factor signaling, and imprinting (IGF2, IGF2R), apoptosis (BAX, BCL2), oxidative stress (MnSOD), metabolism (GLUT1) regulation were observed in cloned embryos. The expression of transcripts in AFSC-NT embryos more closely followed that of the in vitro fertilized embryos compared with AF-NT embryos. It is concluded that AFSCs with a relatively undifferentiated genome may serve as suitable donors which could be reprogrammed more efficiently to reactivate expression of early embryonic genes in buffalo NT.

摘要

由于供体细胞核异常的表观遗传修饰,体细胞核移植胚胎中异常的基因表达可能是观察到的活力下降和发育异常的主要原因。本研究比较了来自成年成纤维细胞(AFs)和羊水干细胞(AFSCs)的水牛核移植(NT)胚胎在4细胞、8至16细胞、桑椹胚和囊胚阶段与发育相关的重要基因表达模式。体外受精胚胎用作对照胚胎。在克隆胚胎中观察到参与转录和多能性(OCT4、STAT3、NANOG)、DNA甲基化(DNMT1、DNMT3A)、组蛋白去乙酰化(HDAC2)、生长因子信号传导和印记(IGF2、IGF2R)、细胞凋亡(BAX、BCL2)、氧化应激(MnSOD)、代谢(GLUT1)调控的基因表达模式发生改变。与AF-NT胚胎相比,AFSC-NT胚胎中转录本的表达更接近体外受精胚胎。得出的结论是,具有相对未分化基因组的AFSCs可能是合适的供体,在水牛NT中可以更有效地被重编程以重新激活早期胚胎基因的表达。

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