Hematopoietic differentiation of human embryonic stem cells progresses through sequential hematoendothelial, primitive, and definitive stages resembling human yolk sac development
Hematopoietic differentiation of human embryonic stem cells progresses through sequential hematoendothelial, primitive, and definitive stages resembling human yolk sac development
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DOI：
10.1182/blood-2004-11-4522
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发表时间：
2005-08-01
期刊：
影响因子：
20.3
通讯作者：
Civin, CI
中科院分区：
文献类型：
--
作者：
Zambidis, ET;Peault, B;Civin, CI
我们阐明的细胞和分子动力学的逐步分化的人胚胎干细胞（hESCs）的原始和永久性红骨髓细胞从人胚状体（hEBs）在无血清克隆形成试验。造血从CD 45 hEB细胞开始，出现半贴壁中胚层-血内皮（MHE）集落，可生成内皮并形成组织化的卵黄囊样结构，进而生成多能原始造血干祖细胞（HSPC）、成红细胞和CD 13（+）CD 45（+）巨噬细胞。第一波造血发生在MHE集落出现之后，主要是以鲜红血红蛋白化、CD 71/CD 325 a（血型糖蛋白A）表达和仅胚胎/胎儿血红蛋白表达为特征的原始红细胞生成。第二波确定型红细胞系爆发形成单位（BFU-e）、红细胞系集落形成单位（CFU-e）、粒细胞-巨噬细胞集落形成细胞（GM-CFC）和多系CFC随后来自hEB祖细胞。这些造血阶段从hEB衍生的细胞自发进行，而不需要在hEB分化期间补充生长因子。对分化中的hEB的基因表达分析显示，造血的起始与SCL/TAL 1、GATA 1、GATA 2、CD 34、CD 31和同源框基因调节因子CDX 4水平的增加相关。这些数据表明，hESC的造血分化以类似于人卵黄囊发育的方式模拟胚胎和定形造血的最早期事件，从而为剖析人类HSPC发生的最早期事件提供了有价值的工具。
We elucidate the cellular and molecular kinetics of the stepwise differentiation of human embryonic stem cells (hESCs) to primitive and definitive erythromyelopoiesis from human embryoid bodies (hEBs) in serum-free clonogenic assays. Hematopoiesis initiates from CD45 hEB cells with emergence of semiadherent mesodermal-hematoendothelial (MHE) colonies that can generate endothelium and form organized, yolk sac-like structures that secondarily generate multipotent primitive hematopoietic stem progenitor cells (HSPCs), erythroblasts, and CD13(+)CD45(+) macrophages. A first wave of hematopoiesis follows MHE colony emergence and is predominated by primitive erythropoiesis characterized by a brilliant red hemoglobinization, CD71/CD325a (glycophorin A) expression, and exclusively embryonic/fetal hemoglobin expression. A second wave of definitive-type erythrold burst-forming units (BFU-e's), erythrold colony-forming units (CFU-e's), granulocyte-macrophage colony-forming cells (GM-CFCs), and multilineage CFCs follows next from hEB progenitors. These stages of hematopoiesis proceed spontaneously from hEB-derived cells without requirement for supplemental growth factors during hEB differentiation. Gene expression analysis of differentiating hEBs revealed that initiation of hematopoiesis correlated with increased levels of SCL/TAL1, GATA1, GATA2, CD34, CD31, and the homeobox gene-regulating factor CDX4 These data indicate that hematopoietic differentiation of hESCs models the earliest events of embryonic and definitive hematopoiesis in a manner resembling human yolk sac development, thus providing a valuable tool for dissecting the earliest events in human HSPC genesis.