一个优化的培养系统以高效地衍生猪扩展的潜在干细胞从植入前胚胎和通过重编程体细胞
Degong Ruan1,2, Yiyi Xuan3, Timothy Theodore Ka Ki Tam3
1Center for Translational Stem Cell Biology, Science Park, Sha Tin, Hong Kong, China.
Nature protocols
|March 21, 2024
概括
研究人员开发了一种强大的方法,从胚胎和重新编程的细胞中创建猪扩展潜在干细胞 (pEPSC). 这些pEPSC对于通过精确的基因组编辑和细胞治疗应用来推进翻译医学和农业生物技术至关重要.
科学领域:
- 干细胞生物学 干细胞生物学
- 翻译医学是一种翻译医学.
- 农业生物技术 农业生物技术
背景情况:
- 由于共享生理学,猪是人类转化医学的宝贵模型.
- 获得猪多能干细胞 (PSC) 对细胞疗法和基因工程至关重要.
- 以前试图隔离猪PSC的尝试面临着挑战.
研究的目的:
- 为猪扩展潜在干细胞 (pEPSCs) 建立一个高度可复制和强大的培养系统.
- 从植入前胚胎和重新编程的体细胞中提取新的pEPSC线条.
- 描述已建立的pEPSC线的属性和基因组编辑能力.
主要方法:
- 为猪EPSC培养物开发一个优化和可重复的协议.
- 从体外受精和体细胞核转移衍生的胚胎中衍生PEPSCs.
- 通过对猪体细胞进行重新编程来衍生PEPSCs.
- 评估pEPSC的自我更新,分化潜力和基因组编辑效率.
主要成果:
- 从不同的胚胎来源成功建立了多个强大的pEPSC系.
- 证明了pEPSC系的长期扩散和稳定特征.
- 在pEPSCs的简单和精确基因组编辑中实现了高效率 (高达100%).
- 在体外和体内确认了胚胎和非胚胎血统的分化潜力.
- 与纤维细胞相比,确定了独特的表观遗传特征,包括较低的H3K27me3水平.
结论:
- 开发的pEPSC培养系统是强大的,可复制的,不需要专业的专业知识.
- pEPSC为推进转化医学和猪农业应用提供了一个强大的平台.
- 在pEPSC中精确编辑猪基因组的能力为研究和生物技术开辟了新的途径.
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