Characterization of two populations of human coronary artery endothelial cells(1).
Characterization of two populations of human coronary artery endothelial cells(1).
复制标题
两个人冠状动脉内皮细胞群的特征(1)。
DOI：
10.1016/j.jss.2004.01.009
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Chen,Changyi
中科院分区：
文献类型：
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作者：
Wu,Huakang;Yao,Qizhi;Lumsden,Alan;Chen,Changyi
内皮细胞的异质性可能影响血管生成、血管愈合和心血管疾病的形成。本研究的目的是鉴定和表征人冠状动脉内皮细胞（HCAECs）的群体、其基因表达水平以及对TNF-α刺激的反应。通过基于尺寸分散分布的流式细胞术分析确定细胞群体。用流式细胞术分析，用适当的门控和抗体荧光标记，研究了有或没有TNF-α（1 ng/ml）刺激16 h的每个群体的基因表达。HCAEC-I和HCAEC-II均显示血管性血友病因子和CD 31阳性染色超过90%，CD 34和CD 4阴性（小于2%）。HCAEC-I的钙粘蛋白-5的表达显著高于HCAEC-II（71.91%vs51.07%）。HCAEC-I还表达更高水平的血管内皮生长因子受体-2（VEGF-R2），（17.35%对0.77%），内皮型一氧化氮合酶（eNOS）（44.64%比2.54%）、VCAM-1（6.08%比1.18%）、E-选择素（18.68%比1.42%）、CXCR 4（61.05%比7.98%）和CCR 5（48.66%比1.97%）。然而，HCAEC-II具有显著高于HCAEC-I的P1 H12（93.07%对58.73%）和ICAM-1（94.37%对58.62%）的表达。在TNF-α刺激后，两个群体的ICAM-1、VCAM-1和E-选择素的表达均显著增加。然而，HCAEC-I，有更高的表达VEGFR-2，eNOS，CXCR 4，CCR-5比HACEC-II后TNF-α stimulation.CONCLUSIONSThese数据表明，商业HCAECs有两个不同的群体具有不同的基因表达模式和不同的反应后，TNF-α刺激。提示冠状动脉内皮细胞的异质性可能具有生物学或病理学意义。
BACKGROUNDHeterogeneity of endothelial cells may affect angiogenesis, vascular healing, and cardiovascular disease formation. The objective of this study was to identify and characterize populations of human coronary artery endothelial cells (HCAECs), their gene expression levels, and response to TNF-α stimulation.MATERIALS AND METHODSCommercial HCAECs were cultured with EGM-2 complete medium. Cell population was determined by flow cytometry analysis based on size-scatter distribution. Gene expression for each population with or without TNF-α (1 ng/ml) stimulation for 16 h was also studied by flow cytometry analysis with proper gating and fluorescence labeling of antibodies.RESULTSTwo distinguished populations, HCAEC-I (small) and HCAEC-II (large), were identified through all passages (from 2 to 10) during cultures. Both HCAEC-I and HCAEC-II showed more than 90% positive staining for both von Willebrand factor and CD31 and were negative for CD34 and CD4 (less than 2%). HCAEC-I had substantially higher expression of cadherin-5 (71.91% versus 51.07%) than HCAEC-II. HCAEC-I also expressed much higher levels of vascular endothelial growth factor receptor-2 (VEGF-R2) (17.35% versus 0.77%), endothelial nitric oxide synthase (eNOS) (44.64% versus 2.54%), VCAM-1 (6.08% versus 1.18%), E-selectin (18.68% versus 1.42%), CXCR4 (61.05% versus 7.98%), and CCR5 (48.66% versus 1.97%) than HCAEC-II. HCAEC-II, however, had substantially higher expression of P1H12 (93.07% versus 58.73%) and ICAM-1 (94.37% versus 58.62%) than HCAEC-I. Following TNF-α stimulation, both populations substantially increased the expression of ICAM-1, VCAM-1, and E-selectin. HCAEC-I, however, had much higher expressions of VEGFR-2, eNOS, CXCR4, and CCR-5 than HACEC-II after TNF-α stimulation.CONCLUSIONSThese data demonstrate that commercial HCAECs have two distinguished populations with different gene expression patterns and different responses following TNF-α stimulation. This study indicates that the heterogeneity of HCAECs may have biologic or pathologic significances in coronary arteries.