Division of Thrombosis and Hemostasis, Department of Internal Medicine, Einthoven Laboratory for Vascular and Regenerative Medicine, Leiden University Medical Center, Leiden, the Netherlands.
Department of Medicine, Queen's University, Kingston, ON, Canada.
J Thromb Haemost. 2020 Oct;18(10):2721-2731. doi: 10.1111/jth.14998. Epub 2020 Aug 18.
Endothelial colony forming cells (ECFCs) derived from peripheral blood can be used to analyze the pathophysiology of vascular diseases ex vivo. However, heterogeneity is observed between ECFC clones and this variability needs to be understood and standardized for ECFCs to be used as a cell model for applications in vascular studies.
Determine reference characteristics of healthy control ECFCs to generate a valid ex vivo model for vascular disease.
Putative ECFCs (n = 47) derived from 21 individual healthy subjects were studied for cell morphology and specific cell characteristics. Clones were analyzed for the production and secretion of von Willebrand factor (VWF), cell proliferation, and the expression of endothelial cell markers.
Based on morphology, clones were categorized into three groups. Group 1 consisted of clones with classic endothelial cell morphology, whereas groups 2 and 3 contained less condensed cells with increasing cell sizes. All clones had comparable endothelial cell surface expression profiles, with low levels of non-endothelial markers. However, a decrease in CD31 and a group-related increase in CD309 and CD45 expression, combined with a decrease in cell proliferation and VWF production and secretion, was observed in clones in group 3 and to a lesser extent in group 2.
We observed group-related variations in endothelial cell characteristics when clones lacked the classic endothelial cell morphology. Despite this variation, clones in all groups expressed endothelial cell surface markers. Provided that clones with similar characteristics are compared, we believe ECFCs are a valid ex vivo model to study vascular disease.
外周血来源的内皮祖细胞（ECFC）可用于体外分析血管疾病的病理生理学。然而，ECFC 克隆之间存在异质性，需要了解这种变异性，并将其标准化，以便将 ECFC 用作血管研究应用的细胞模型。
确定健康对照 ECFC 的参考特征，以生成用于血管疾病的有效体外模型。
研究了来自 21 名健康个体的 47 个推定 ECFC，以研究细胞形态和特定细胞特征。对克隆进行 von Willebrand 因子（VWF）的产生和分泌、细胞增殖和内皮细胞标志物的表达分析。
根据形态，将克隆分为三组。第 1 组由具有经典内皮细胞形态的克隆组成，而第 2 组和第 3 组包含细胞大小增大、凝聚性降低的细胞。所有克隆均具有相似的内皮细胞表面表达谱，非内皮标志物水平较低。然而，在第 3 组和在较小程度上在第 2 组中观察到 CD31 减少，以及与群组相关的 CD309 和 CD45 表达增加，同时细胞增殖和 VWF 产生和分泌减少。
当克隆缺乏经典的内皮细胞形态时，我们观察到与群组相关的内皮细胞特征变化。尽管存在这种变化，但所有群组的克隆均表达内皮细胞表面标志物。只要比较具有相似特征的克隆，我们就相信 ECFC 是研究血管疾病的有效体外模型。
