Directed differentiation and mass cultivation of pure erythroid progenitors from mouse embryonic stem cells
Directed differentiation and mass cultivation of pure erythroid progenitors from mouse embryonic stem cells
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DOI：
10.1182/blood-2004-02-0570
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发表时间：
2004-09-15
期刊：
影响因子：
20.3
通讯作者：
Beug, H
中科院分区：
文献类型：
--
作者：
Carotta, S;Pilat, S;Beug, H
分化的胚胎干细胞是造血祖细胞的重要来源，可用于基础研究和临床应用。除了它们在集落测定中的表征之外，还存在用于培养淋巴样、髓样和红细胞的方案。然而，除了肥大细胞之外，如果没有外源基因过度表达引起的永生化，从ES细胞长期扩增纯造血祖细胞是不可能的。在这里，我们首次描述了一种有效而简单的策略，使用无血清培养基加上重组细胞因子和激素，从小鼠ES细胞（ES-EPs）中大量培养纯的未成熟红系祖细胞。ES-EP代表长寿命的、成年的、永久性红系祖细胞，其类似于在应激红细胞生成期间在体内扩增的未成熟红系细胞。当暴露于终末分化条件时，ES-EP分化为成熟的去核红细胞。重要的是，注射到小鼠中的ES-EP没有表现出致瘤潜力，但分化成正常的红细胞。几乎无限的细胞供应和确定的培养条件使我们的系统成为分析影响红系祖细胞增殖和成熟的因素的有价值的工具。此外，该系统允许使用野生型（wt）和遗传修饰的ES细胞在红系细胞增殖和分化过程中的详细表征。(C)2004年，美国血液学会。
Differentiating embryonic stem (ES) cells are an increasingly important source of hematopoletic progenitors, useful for both basic research and clinical applications. Besides their characterization in colony assays, protocols exist for the cultivation of lymphoid, myelold, and erythrold cells. With the possible exception of mast cells, however, long-term expansion of pure hematopoietic progenitors from ES cells has not been possible without immortalization caused by overexpression of exogenous genes. Here, we describe for the first time an efficient yet easy strategy to generate mass cultures of pure, immature erythroid progenitors from mouse ES cells (ES-EPs), using serum-free medium plus recombinant cytolkines and hormones. ES-EPs represent long-lived, adult, definitive erythroid progenitors that resemble immature erythroid cells expanding in vivo during stress erythropoiesis. When exposed to terminal differentiation conditions, ES-EPs differentiated into mature, enucleated erythrocytes. Importantly, ES-EPs injected into mice did not exhibit tumorigenic potential but differentiated into normal erythrocytes. Both the virtually unlimited supply of cells and the defined culture conditions render our system a valuable tool for the analysis of factors influencing proliferation and maturation of erythroid progenitors. In addition, the system allows detailed characterization of processes during erythroid proliferation and differentiation using wild-type (wt) and genetically modified ES cells. (C) 2004 by The American Society of Hematology.