In vitro hematopoietic and endothelial cell development from cells expressing TEK receptor in murine aorta-gonad-mesonephros region
In vitro hematopoietic and endothelial cell development from cells expressing TEK receptor in murine aorta-gonad-mesonephros region
复制标题
DOI：
10.1182/blood.v93.5.1549.405k25_1549_1556
复制
发表时间：
1999-03-01
期刊：
影响因子：
20.3
通讯作者：
Suda, T
中科院分区：
文献类型：
--
作者：
Hamaguchi, I;Huang, XL;Suda, T
最近的研究表明，长期再生的造血干细胞(HSCs)首先出现在主动脉-性腺-中肾(AGM)区。免疫组织化学研究表明，在AGM区存在TEK+细胞。约5%的AGM细胞为TEK+，其中大部分为CD34(+)和c-Kit(+)。然后，我们利用缺乏巨噬细胞集落刺激因子(M-CSF)的基质细胞系OP9建立了AGM细胞的共培养体系。利用该系统，我们发现AGM细胞在性交后10.5天(DPC)分化并增殖为造血细胞和内皮细胞。增殖的造血细胞在培养(CFU-C)和脾(CFU-S)中均含有大量的集落形成细胞。在10.5dPC的原代AGM细胞中，分离的TEK+AGM细胞在OP9基质层产生造血细胞和血小板内皮细胞黏附分子(PECAM)-1(+)内皮细胞，而TEK-细胞不产生。当在AGM的单细胞培养中加入TEK的配体Angiopoietin-1时，在造血细胞集落生长的孔中检测到内皮细胞的生长。虽然发生率仍然很低(1/135)，但我们发现，使用单细胞沉积系统，单个TEK+细胞同时产生造血细胞和内皮细胞。这种体外共培养体系表明，原代AGM细胞的TEK+组分是血管母细胞的候选细胞，可以分化为造血细胞和内皮细胞。(C)1999年由美国血液病学会主办。
Recent studies have shown that long-term repopulating hematopoietic stem cells (HSCs) first appear in the aorta-gonad-mesonephros (AGM) region. Our immunohistochemistry study showed that TEK+ cells existed in the AGM region. Approximately 5% of AGM cells were TEK+, and most of these were CD34(+) and c-Kit(+). We then established a coculture system of AGM cells using a stromal cell line, OP9, which is deficient in macrophage colony-stimulating factor (M-CSF). With this system, we showed that AGM cells at 10.5 days postcoitum (dpc) differentiated and proliferated into both hematopoietic and endothelial cells. Proliferating hematopoietic cells contained a significant number of colony-forming cells in culture (CFU-C) and in spleen (CFU-S). Among primary AGM cells at 10.5 dpc, sorted TEK+ AGM cells generated hematopoietic cells and platelet endothelial cell adhesion molecule (PECAM)-1(+) endothelial cells on the OP9 stromal layer, while TEK- cells did not. When a ligand for TEK, angiopoietin-1, was added to the single-cell culture of AGM, endothelial cell growth was detected in the wells where hematopoietic colonies grew. Although the incidence was still low (1/135), we showed that single TEK+ cells generated hematopoietic cells and endothelial cells simultaneously, using a single-cell deposition system. This in vitro coculture system shows that the TEK+ fraction of primary AGM cells is a candidate for hemangioblasts, which can differentiate into both hematopoietic cells and endothelial cells. (C) 1999 by The American Society of Hematology.