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细胞提取物通过改变DNA甲基化和多能性基因表达促进核重编程。

Cellular extract facilitates nuclear reprogramming by altering DNA methylation and pluripotency gene expression.

作者信息

Xiong Xian-Rong, Lan Dao-Liang, Li Jian, Zi Xiang-Dong, Ma Li, Wang Yong

机构信息

1 College of Life Science and Technology, Southwest University for Nationalities , Chengdu, Sichuan, 610041, China .

出版信息

Cell Reprogram. 2014 Jun;16(3):215-22. doi: 10.1089/cell.2013.0078. Epub 2014 Apr 16.

Abstract

The functional reprogramming of a differentiated cell to a pluripotent state presents potential beneficial applications in disease mechanisms and regenerative medicine. Epigenetic modifications enable differentiated cells to perpetuate molecular memory to retain their identity. Therefore, the aim of this study was to investigate the reprogramming modification of yak fibroblast cells that were permeabilized and incubated in the extracts of mesenchymal stem cells derived from mice adipose tissue [adipose-derived stem cells (ADSCs)]. According to the results, the treatment of ADSC extracts promoted colony formation. Moreover, pluripotent gene expression was associated with the loss of repressive histone modifications and increased global demethylation. The genes Col1a1 and Col1a2, which are typically found in differentiated cells only, demonstrated decreased expression and increased methylation in the 5'-flanking regulatory regions. Moreover, yak fibroblast cells that were exposed to ADSC extracts resulted in significantly different eight-cell and blastocyst formation rates of cloned embryos compared with their untreated counterparts. This investigation provides the first evidence that nuclear reprogramming of yak fibroblast cells is modified after the ADSC extract treatment. This research also presents a methodology for studying the dedifferentiation of somatic cells that can potentially lead to an efficient way of reprogramming somatic cells toward a pluripotent state without genetic alteration.

摘要

将分化细胞重编程为多能状态在疾病机制和再生医学中具有潜在的有益应用。表观遗传修饰使分化细胞能够延续分子记忆以保持其身份。因此,本研究的目的是探讨牦牛成纤维细胞在经小鼠脂肪组织来源的间充质干细胞提取物(脂肪来源干细胞,ADSCs)通透处理并孵育后的重编程修饰。结果显示,ADSC提取物处理促进了集落形成。此外,多能基因表达与抑制性组蛋白修饰的丧失和整体去甲基化增加有关。通常仅在分化细胞中发现的Col1a1和Col1a2基因,在5'侧翼调控区域的表达降低且甲基化增加。此外,与未处理的牦牛成纤维细胞相比,暴露于ADSC提取物的牦牛成纤维细胞导致克隆胚胎的八细胞和囊胚形成率有显著差异。本研究首次证明了ADSC提取物处理后牦牛成纤维细胞的核重编程发生了改变。该研究还提出了一种研究体细胞去分化的方法,这可能会导致一种在不进行基因改造的情况下将体细胞高效重编程为多能状态的方法。

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