Scalable In Vitro Production of Defined Mouse Erythroblasts
Scalable In Vitro Production of Defined Mouse Erythroblasts
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确定的小鼠成红细胞的可扩展体外生产
DOI：
10.1101/2020.11.10.376749
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Francis H
中科院分区：
文献类型：
--
作者：
Francis H
小鼠胚胎干细胞（mESC）可以在体外模拟红细胞生成的过程，即多能细胞经过谱系特化、分化和成熟产生红系细胞。尽管该系统可用于鉴定特定的祖细胞和前体细胞，但尚未被充分利用作为分析红细胞生成的细胞来源。在这里，我们建立了一个协议，其中特征成红细胞可以分离在一个可扩展的方式从分化的胚状体（EB）。使用转录和表观遗传学分析，我们证明，该系统忠实地重演正常的原始红细胞生成，并完全再现从小鼠模型获得的原代细胞中看到的天然和工程突变的影响。我们预计，该系统是在减少时间和成本的产生和维护鼠标线在一些研究方案的巨大价值。
Mouse embryonic stem cells (mESCs) can be manipulatedin vitroto recapitulate the process of erythropoiesis, during which multipotent cells undergo lineage specification, differentiation and maturation to produce erythroid cells. Although useful for identifying specific progenitors and precursors, this system has not been fully exploited as a source of cells to analyse erythropoiesis. Here, we establish a protocol in which characterised erythroblasts can be isolated in a scalable manner from differentiated embryoid bodies (EBs). Using transcriptional and epigenetic analysis, we demonstrate that this system faithfully recapitulates normal primitive erythropoiesis and fully reproduces the effects of natural and engineered mutations seen in primary cells obtained from mouse models. We anticipate this system to be of great value in reducing the time and costs of generating and maintaining mouse lines in a number of research scenarios.