基底状态的雌性ES细胞的衍生,维护配体衍生的DNA甲基化
Masaki Yagi1, Satoshi Kishigami2, Akito Tanaka1
1Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Nature
|July 27, 2017
概括
在小鼠胚胎干细胞 (ES细胞) 衍生中使用的两个小分子 (2i) 消除了基因组印记. 虽然这些细胞可以分化,但它们显示出发育缺陷,突出显示出需要改进的衍生方法.
科学领域:
- 发育生物学
- 表观遗传学
- 干细胞生物学
背景情况:
- 两种小分子 (2i) 和白血病抑制因子 (LIF) 增强胚胎干细胞 (ES细胞) 的衍生,并促进动物的基态多能性.
- 用2i/LIF衍生雌性小鼠ES细胞导致广泛的DNA低甲基化和基因组印记的删除.
研究的目的:
- 研究2i/LIF对雌性小鼠ES细胞的DNA甲基化,印记和发育潜力的影响.
- 识别能够保持多能性的衍生条件,同时保持表观遗传标记和发育能力.
主要方法:
- 在存在2i/LIF的雌性小鼠ES细胞的衍生.
- 包括印记控制区域 (ICR) 在内的DNA甲基化分析.
- 测定四平体胚胎补充和核移植以评估发育潜力.
主要成果:
- 2i/LIF治疗导致全局DNA低甲基化和雌性ES细胞中的基因组印记的大规模删除.
- 尽管低甲基化,早期的2i/LIF ES细胞可以分化为体细胞,需要新的DNA甲基化.
- 然而,大多数ICR仍未在分化细胞中甲基化,而2i/ LIF ES细胞显示胚胎和胎盘发育受损.
- 长时间培养导致ICR脱甲基,不论培养条件如何.
- 特定的衍生条件被确定为产生具有2i/LIF类转录特征的雌性ES细胞,但保留DNA甲基化和发育潜力.
结论:
- 使用2i/LIF用于雌性ES细胞衍生导致表观遗传异常,包括印记删除,损害发育潜力.
- 维护配体衍生的DNA甲基化对于维持ES细胞的发育能力至关重要.
- 这项研究提供了对提取女性ES细胞的见解,这些细胞模仿了植入前胚胎的内部细胞质量,为发育研究提供了更强大的模型.
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