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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

机译:在成年成纤维细胞和羊水衍生干细胞衍生的水牛手工克隆胚胎中植入前阶段重要发育基因表达谱的比较研究

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摘要

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.
机译:由于供体核的表观遗传学异常修饰,导致体细胞核移植胚胎中的基因表达异常,可能是导致观察到的活力降低和发育异常的主要原因。本研究比较了成年成纤维细胞(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ula和胚泡阶段的重要发育基因表达模式。体外受精的胚胎用作对照胚胎。涉及转录和多能性(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迹(IGF2,IGF2R),凋亡(BAX, BCL2),氧化应激(MnSOD),代谢(GLUT1)调节在克隆的胚胎中观察到。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。结论是,具有相对未分化的基因组的AFSCs可以作为合适的供体,可以更有效地重新编程以重新激活水牛NT中早期胚胎基因的表达。

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