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多能性相关基因的表达高度依赖于三氮唑乙酸辅助的猪间充质干细胞的表观基因组调节,这些细胞经过凋亡分析,随后用于生成克隆胚胎。

Expression of pluripotency-related genes is highly dependent on trichostatin A-assisted epigenomic modulation of porcine mesenchymal stem cells analysed for apoptosis and subsequently used for generating cloned embryos.

机构信息

Department of Reproductive Biotechnology and Cryoconservation, National Research Institute of Animal Production, Balice n. Kraków, Poland.

Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Poznań, Poland.

出版信息

Anim Sci J. 2019 Sep;90(9):1127-1141. doi: 10.1111/asj.13260. Epub 2019 Jul 12.

Abstract

The present study sought to examine whether trichostatin A (TSA)-assisted epigenetic transformation of porcine bone marrow (BM)-derived mesenchymal stem cells (BM-MSCs) affects the transcriptional activities of pluripotency-related genes (Oct4, Nanog, c-Myc, Sox2 and Rex1), multipotent stemness-related gene (Nestin) and anti-apoptotic/anti-senescence-related gene (Survivin). Epigenetically transformed or non-transformed BM-MSCs that had been transcriptionally profiled by qRT-PCR and had been analysed for different stages of apoptosis progression provided a source of nuclear donor cells for the in vitro production of cloned pig embryos. TSA-mediated epigenomic modulation has been found to enhance the multipotency extent, stemness and intracellular anti-ageing properties of porcine BM-MSCs. This has been confirmed by the relative abundances for Nanog, c-Myc Rex1, Sox2 and Survivin mRNAs in TSA-exposed BM-MSCs that turned out to be significantly higher than those of TSA-unexposed BM-MSCs. Additionally, TSA-assisted epigenomic modulation of BM-MSCs did not impact the caspase-8 activity, Bax protein expression and the incidence of TUNEL-positive cells. In conclusion, the considerably elevated quantitative profiles of Sox2, Rex1, c-Myc, Nanog and Survivin mRNA transcripts seem to trigger improved reprogrammability of TSA-treated BM-MSC nuclei in cloned pig embryos that thereby displayed remarkably increased blastocyst formation rates as compared to those noticed for embryos derived from TSA-untreated BM-MSCs.

摘要

本研究旨在探讨曲古抑菌素 A(TSA)辅助的猪骨髓(BM)来源间充质干细胞(BM-MSCs)的表观遗传转化是否影响多能性相关基因(Oct4、Nanog、c-Myc、Sox2 和 Rex1)、多能性相关基因(Nestin)和抗凋亡/抗衰老相关基因(Survivin)的转录活性。通过 qRT-PCR 进行转录谱分析并分析不同凋亡进展阶段的表观遗传转化或未转化的 BM-MSCs 为体外生产克隆猪胚胎提供了核供体细胞来源。已发现 TSA 介导的表观基因组调节可增强猪 BM-MSCs 的多能性程度、干细胞特性和细胞内抗老化特性。这一点已通过 TSA 暴露的 BM-MSCs 中 Nanog、c-Myc Rex1、Sox2 和 Survivin mRNA 的相对丰度得到证实,其明显高于 TSA 未暴露的 BM-MSCs。此外,TSA 辅助的 BM-MSCs 表观基因组调节不会影响 caspase-8 活性、Bax 蛋白表达和 TUNEL 阳性细胞的发生率。总之,Sox2、Rex1、c-Myc、Nanog 和 Survivin mRNA 转录本的数量明显增加似乎触发了 TSA 处理的 BM-MSC 核在克隆猪胚胎中的重编程能力提高,从而与源自 TSA 未处理的 BM-MSCs 的胚胎相比,胚胎的囊胚形成率显著增加。

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