Large-scale production of embryonic red blood cells from human embryonic stem cells
Large-scale production of embryonic red blood cells from human embryonic stem cells
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DOI：
10.1016/j.exphem.2006.07.003
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发表时间：
2006-12-01
影响因子：
2.6
通讯作者：
Bouhassira, Eric E.
中科院分区：
文献类型：
--
作者：
Olivier, Emmanuel N.;Qiu, Caihong;Bouhassira, Eric E.
目标。目的：建立一种从人胚胎干细胞中培养大量红系细胞的方法。材料与方法。将人H1胚胎干细胞与人胎儿肝细胞系共培养分化为造血细胞，并采用三步法在体外液体培养中扩增得到cd34阳性细胞。然后用光镜和流式细胞术分析产生的红细胞。采用定量逆转录酶聚合酶链反应和高效液相色谱法对珠蛋白的表达进行了表征。人胚胎干细胞产生的cd34阳性细胞在液体培养中能有效地分化为红系细胞，细胞数量增加5000倍以上。所产生的红细胞与早期人类胚胎卵黄囊中存在的原始红细胞相似，不去核。它们是完全血红蛋白化的，表达胚胎和胎儿的混合球蛋白，但不表达-球蛋白。我们已经开发了一种实验方案，从未分化的人类胚胎干细胞开始生产大量原始红细胞。由于人类最早的红细胞，即有核的原始红母细胞，在这个发育阶段的实验材料很难获得，所以还没有很好地表征，这个系统应该证明对回答关于这些细胞的生物学的一些实验问题是有用的。此外，从人类胚胎干细胞中生产成熟红细胞具有巨大的潜在实际重要性，因为它最终可能成为输血细胞的替代来源。(c) 2006年国际实验血液学学会。Elsevier Inc.出版。
Objective. To develop a method to produce in culture large number of erythroid cells from human embryonic stem cells.Materials and Methods. Human H1 embryonic stem cells were differentiated into hematopoietic cells by coculture with a human fetal liver cell line, and the resulting CD34-positive cells were expanded in vitro in liquid culture using a three-step method. The erythroid cells produced were then analyzed by light microscopy and flow cytometry. Globin expression was characterized by quantitative reverse-transcriptase polymerase chain reaction and by high-performance liquid chromatography.Results. CD34-positive cells produced from human embryonic stem cells could be efficiently differentiated into erythroid cells in liquid culture leading to a more than 5000-fold increase in cell number. The erythroid cells produced are similar to primitive erythroid cells present in the yolk sac of early human embryos and did not enucleate. They are fully hemoglobinized and express a mixture of embryonic and fetal globins but no beta-globin.Conclusions. We have developed an experimental protocol to produce large numbers of primitive erythroid cells starting from undifferentiated human embryonic stem cells. As the earliest human erythroid cells, the nucleated primitive erythroblasts, are not very well characterized because experimental material at this stage of development is very difficult to obtain, this system should prove useful to answer a number of experimental questions regarding the biology of these cells. In addition, production of mature red blood cells from human embryonic stem cells is of great potential practical importance because it could eventually become an alternate source of cell for transfusion. (c) 2006 International Society for Experimental Hematology. Published by Elsevier Inc.