Identification and characterization of hemoangiogenic progenitors during cynomolgus monkey embryonic stem cell differentiation
Identification and characterization of hemoangiogenic progenitors during cynomolgus monkey embryonic stem cell differentiation
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DOI：
10.1634/stemcells.2005-0165
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发表时间：
2006-05-01
期刊：
影响因子：
5.2
通讯作者：
Nakahata, Tatsutoshi
中科院分区：
文献类型：
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作者：
Umeda, Katsutsugu;Heike, Toshio;Nakahata, Tatsutoshi
我们从非人灵长类动物胚胎干细胞（ES）中鉴定出了具有分化为造血和内皮细胞的潜能的中期祖细胞。连续荧光激活细胞分选和免疫染色分析表明，当ES细胞在OP 9共培养系统中培养时，两种谱系在两种具有血血管生成祖细胞的细胞组分，即血管内皮生长因子受体（VEGFR）-2（high）CD 34（-）和VEGFR-2（high）CD 34（+）细胞出现后发育。外源性血管内皮生长因子以剂量依赖性方式增加VEGFR-2高表达细胞的比例，特别是VEGFR-2高表达的CD 34（+）细胞的比例。尽管任一VEGFR-2（高）细胞群体均可分化为原始和永久造血细胞（HC）以及内皮细胞（EC），但VEGFR-2（高）CD 34+细胞具有更大的血血管生成潜力。这两种谱系在单细胞水平上均由VEGFR-2（高）CD 34-或VEGFR-2（高）CD 34+前体发育而来，这强烈支持这些细胞组分中存在成血管细胞。因此，该培养系统允许分化成HC和EC谱系，以通过表面标记物来定义。这些观察结果将有助于对人类早期发育过程和再生治疗的进一步研究。
We identified intermediate-stage progenitor cells that have the potential to differentiate into hematopoietic and endothelial lineages from nonhuman primate embryonic stem (ES) cells. Sequential fluorescence-activated cell sorting and immunostaining analyses showed that when ES cells were cultured in an OP9 coculture system, both lineages developed after the emergence of two hemoangiogenic progenitor-bearing cell fractions, namely, vascular endothelial growth factor receptor (VEGFR)-2(high) CD34(-) and VEGFR-2(high) CD34(+) cells. Exogenous vascular endothelial growth factor increased the proportion of VEGFR-2 high cells, particularly that of VEGFR-2 high CD34(+) cells, in a dose-dependent manner. Although either population of VEGFR-2(high) cells could differentiate into primitive and definitive hematopoietic cells (HCs), as well as endothelial cells (ECs), the VEGFR-2(high) CD34+ cells had greater hemoangiogenic potential. Both lineages developed from VEGFR-2(high) CD34- or VEGFR-2(high) CD34+ precursor at the single-cell level, which strongly supports the existence of hemangioblasts in these cell fractions. Thus, this culture system allows differentiation into the HC and EC lineages to be defined by surface markers. These observations should facilitate further studies both on early developmental processes and on regeneration therapies in human.