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一种利用全机械方法从废弃胚胎中建立人胚胎干细胞系 HN4 的方法。

A whole-mechanical method to establish human embryonic stem cell line HN4 from discarded embryos.

机构信息

Reproductive Medical Center of Hainan Province, Hainan Medical College, 570102, Haikou, China,

出版信息

Cytotechnology. 2010 Dec;62(6):509-18. doi: 10.1007/s10616-010-9311-z. Epub 2010 Oct 26.

Abstract

Since the first human embryonic stem cell (hESC) line was generated by Thomson et al. (in Science 282:1145-1147, 1998), hundreds of hESC lines have been reported by different labs, providing resources for basic research and regenerative medicine as well. However it has been widely recognized that hESC lines varied on their properties, in terms of gene expression profile, epigenetic modify profile, and differentiation tendency. Generation of more hESC lines will largely enhance our knowledge of hESCs innate character. In this current work, we reported the generation of HN4, a hESC line derived from grade III IVF human embryo by using a mixture of human foreskin fibroblast (HFF) and mouse embryonic fibroblast (MEF) as feeder layers, and a whole-mechanical method in inner cell mass (ICM) isolation. HN4 satisfied the criteria of hESCs pluripotency, with high expression of hESC surface markers (SSEA-3, SSEA-4, TRA-1-60, TRA-1-81), transcription factors (OCT-4, NANOG, REX-1), and alkaline phosphatase. It is able to differentiate to three germ layer derivatives when cultured in vitro, or in teratoma formation. Moreover, it displayed promising potential in neural differentiation under a proper culture condition, suggesting the advantage of HN4 in further investigation. Additionally, the whole-mechanical protocol for ICM isolation facilitates hESC line generation for its ease to handle.

摘要

自从 Thomson 等人(在《科学》282:1145-1147,1998 年)首次生成人类胚胎干细胞(hESC)系以来,不同实验室已经报道了数百种 hESC 系,为基础研究和再生医学提供了资源。然而,人们广泛认识到,hESC 系在基因表达谱、表观遗传修饰谱和分化倾向等方面存在差异。生成更多的 hESC 系将大大增强我们对 hESC 固有特性的了解。在本研究中,我们报道了 HN4 的生成,这是一种通过使用人包皮成纤维细胞(HFF)和小鼠胚胎成纤维细胞(MEF)混合物作为饲养层,以及全机械方法分离内细胞团(ICM),从 III 级 IVF 人类胚胎中衍生的 hESC 系。HN4 满足 hESC 多能性的标准,具有高表达的 hESC 表面标志物(SSEA-3、SSEA-4、TRA-1-60、TRA-1-81)、转录因子(OCT-4、NANOG、REX-1)和碱性磷酸酶。当在体外培养或在畸胎瘤形成中时,它能够分化为三个胚层衍生物。此外,在适当的培养条件下,它显示出有前途的神经分化潜力,表明 HN4 在进一步研究中的优势。此外,ICM 分离的全机械方案因其易于操作而有利于 hESC 系的生成。

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