A Boost of BMP4 Accelerates the Commitment of Human Embryonic Stem Cells to the Endothelial Lineage
A Boost of BMP4 Accelerates the Commitment of Human Embryonic Stem Cells to the Endothelial Lineage
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DOI：
10.1002/stem.100
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发表时间：
2009-08-01
期刊：
影响因子：
5.2
通讯作者：
Uzan, Georges
中科院分区：
文献类型：
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作者：
Goldman, Orit;Feraud, Olivier;Uzan, Georges
人胚胎干细胞（human embryonic stem cells，hESC）分化过程中产生的类胚体（Embryoid bodies，EBs）含有血管样结构，提示中胚层祖细胞向内皮细胞的定向分化是自发发生的。我们发现，骨形态发生蛋白4（BMP 4），中胚层的诱导剂，加速了CD 133/激酶插入结构域包含受体（KDR）和CD 144/KDR的峰值表达。因为CD 133（+）KDR（+）群体可以代表内皮祖细胞，所以我们在第7天将其分选并在内皮培养基中培养。然而，这些细胞不能分化成内皮细胞。在标准条件下，在第12天，CD 144（+）KDR（+）群体占总细胞的10%。在培养中，这些细胞如果被分选，则产生具有内皮细胞典型形态的同质群体，并表达内皮标志物。如通过不同的体外测定所评估的，这些源自第12天分选群体的内皮细胞是功能性的。当EB被BMP 4刺激时，CD 144（+）KDR（+）峰移动到第7天。然而，这些细胞中的大多数是CD 31（-），在培养中变成CD 31（+）。然后，它们表达冯维勒布兰德因子并具有功能。这表明，最初，BMP 4增强的第7天，CD 144（+）KDR（+）CD 31（-）群体代表在培养中分化为成熟内皮细胞的未成熟内皮细胞。在EB分化过程中，hESC不成熟的标志物OCT 3/4的表达降低，在BMP 4诱导后降低得更快。我们还表明，BMP 4抑制GATA 2和RUNX 1，两个转录因子参与成血管细胞形成，在第7天和第12天的全球表达。干细胞2009;27：1750-1759
Embryoid bodies (EBs) generated during differentiation of human embryonic stem cells (hESCs) contain vascular-like structures, suggesting that commitment of mesoderm progenitors into endothelial cells occurs spontaneously. We showed that bone morphogenetic protein 4 (BMP4), an inducer of mesoderm, accelerates the peak expression of CD133/kinase insert domain-containing receptor (KDR) and CD144/KDR. Because the CD133(+)KDR(+) population could represent endothelial progenitors, we sorted them at day 7 and cultured them in endothelial medium. These cells were, however, unable to differentiate into endothelial cells. Under standard conditions, the CD144(+)KDR(+) population represents up to 10% of the total cells at day 12. In culture, these cells, if sorted, give rise to a homogeneous population with a morphology typical of endothelial cells and express endothelial markers. These endothelial cells derived from the day 12 sorted population were functional, as assessed by different in vitro assays. When EBs were stimulated by BMP4, the CD144(+)KDR(+) peak was shifted to day 7. Most of these cells, however, were CD31(-), becoming CD31(+) in culture. They then expressed von Willebrand factor and were functional. This suggests that, initially, the BMP4-boosted day 7, CD144(+)KDR(+)CD31(-) population represents immature endothelial cells that differentiate into mature endothelial cells in culture. The expression of OCT3/4, a marker of immaturity for hESCs decreases during EB differentiation, decreasing faster following BMP4 induction. We also show that BMP4 inhibits the global expression of GATA2 and RUNX1, two transcription factors involved in hemangioblast formation, at day 7 and day 12. STEM CELLS 2009;27:1750-1759