Different differentiation kinetics of vascular progenitor cells in primate and mouse embryonic stem cells
Different differentiation kinetics of vascular progenitor cells in primate and mouse embryonic stem cells
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DOI：
10.1161/01.cir.0000070022.78747.1b
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发表时间：
2003-04-29
期刊：
影响因子：
37.8
通讯作者：
Nakao, K
中科院分区：
文献类型：
--
作者：
Sone, M;Itoh, H;Nakao, K
背景-我们证明了来自小鼠胚胎干细胞的血管内皮生长因子受体2（VEGF-R2）阳性细胞可以分化为内皮细胞和壁细胞，以满足血管祖细胞（VPC）的要求。在这里，我们研究是否VPC发生在灵长类动物ES细胞和研究的差异，在VPC分化动力学之间的灵长类动物和小鼠ES cells.Methods和Results-In对比小鼠ES细胞，未分化的猴ES细胞表达VEGF-R2。通过将这些未分化的ES细胞在OP 9饲养层上培养4天，VEGF-R2表达消失，然后在分化8天后重新出现。然后我们通过流式细胞术分选分离这些VEGF-R2阳性和血管内皮钙粘蛋白（VECadherin）阴性的细胞。将这些VEGF-R2（+）VE钙粘蛋白（-）细胞在OP 9饲养层上再培养5天，导致出现血小板内皮细胞粘附分子-1（PECAM 1）阳性、VE钙粘蛋白阳性、内皮一氧化氮合酶（eNOS）阳性内皮细胞。在胶原IV包被的培养皿中，在血清的存在下，这些细胞分化成平滑肌肌动蛋白（SMA）阳性和钙调蛋白阳性的壁细胞（周细胞或血管平滑肌细胞）。在胶原IV包被的培养皿中加入50 ng/mL VEGF导致SMA（+）细胞包围PECAM 1（+）细胞的出现。此外，这些分化的VEGF-R2（+）细胞可以在三维culture.Conclusion-Our研究结果表明，来自灵长类动物和小鼠ES细胞的VPCs的分化动力学是不同的。分化的VEGF-R2（+）VECadherin（-）细胞可以作为灵长类动物的VPC。为了探索VPCs在血管再生中的临床应用潜力，对灵长类ES细胞的研究是必不可少的。
Background-We demonstrated that vascular endothelial growth factor receptor 2 (VEGF-R2)-positive cells derived from mouse embryonic stem (ES) cells can differentiate into both endothelial cells and mural cells to suffice as vascular progenitor cells (VPCs). Here we examined whether VPCs occur in primate ES cells and investigated the differences in VPC differentiation kinetics between primate and mouse ES cells.Methods and Results-In contrast to mouse ES cells, undifferentiated monkey ES cells expressed VEGF-R2. By culturing these undifferentiated ES cells for 4 days on OP9 feeder layer, VEGF-R2 expression disappeared, and then reappeared after 8 days of differentiation. We then isolated these VEGF-R2 -positive and vascular endothelial cadherin (VEcadherin)-negative cells by flow cytometry sorting. Additional 5-day reculture of these VEGF-R2(+) VEcadherin(-) cells on OP9 feeder layer resulted in the appearance of platelet endothelial cell adhesion molecule-1 (PECAM1)-positive, VEcadherin- positive, endothelial nitric oxide synthase (eNOS)-positive endothelial cells. On a collagen IV-coated dish in the presence of serum, these cells differentiated into smooth muscle actin (SMA)-positive and calponin-positive mural cells (pericytes or vascular smooth muscle cells). Addition of 50ng/mL VEGF to the culture on a collagen IV-coated dish resulted in the appearance of PECAM1(+) cells surrounded by SMA(+) cells. In addition, these differentiated VEGF-R2(+) cells can form tube-like structures in a 3-dimensional culture.Conclusion-Our findings indicate that differentiation kinetics of VPCs derived from primate and mouse ES cells were different. Differentiated VEGF-R2(+) VEcadherin(-) cells can act as VPCs in primates. To seek the clinical potential of VPCs for vascular regeneration, investigations of primate ES cells are indispensable.