用于研究 HDL 功能的永生化人真皮微血管内皮细胞（HMEC-1）的特征。
Characterization of immortalized human dermal microvascular endothelial cells (HMEC-1) for the study of HDL functionality.
机构信息
Molecular Biology Department, Instituto Nacional de Cardiología "Ignacio Chávez", Juan Badiano 1, Sección XVI, 14080, Mexico City, Mexico.
Physiology Departments, Instituto Nacional de Cardiología "Ignacio Chávez", Mexico City, Mexico.
出版信息
Lipids Health Dis. 2018 Mar 9;17(1):44. doi: 10.1186/s12944-018-0695-7.
BACKGROUND
Primary cultures endothelial cells have been used as models of endothelial related diseases such atherosclerosis. Biological behavior of primary cultures is donor-dependent and data could not be easily reproducible; endothelial cell lines are emerging options, particularly, human dermal microvascular endothelial cells (HMEC-1), that should be validated to substitute primary cultures for the study of HDL functions.
METHODS
Morphology, size and granularity of cells were assessed by phase contrast microscopy and flow cytometry of HMEC-1. The adhesion molecules, ICAM-1and VCAM-1 after TNF-α stimulation, and endothelial markers CD105 endoglin, as well as HDL receptor SR-BI were determined by flow cytometry. Internalization of HDL protein was demonstrated by confocal microscopy using HDL labeled with Alexa Fluor 488. HUVECs were used as reference to compared the characteristics with HMEC-1.
RESULTS
HMEC-1 and HUVEC had similar morphologies, size and granularity. HMEC-1 expressed endothelial markers as HUVECs, as well as functional SR-B1 receptor since the cell line was able to internalize HDL particles. HMEC-1 effectively increased ICAM-1 and VCAM-1 expression after TNF-α stimulation. HUVECs showed more sensibility to TNF-α stimulus but the range of ICAM-1 and VCAM-1 expression was less homogeneous than in HMEC-1, probably due to biological variation of the former. Finally, the expression of adhesion molecules in HMEC-1 was attenuated by co-incubation with HDL.
CONCLUSION
HMEC-1 possess characteristics of endothelial cells, similar to HUVECs, being a cell line suitable to evaluate the functionality of HDL vis-à-vis the endothelium.
背景
原代培养的内皮细胞已被用于动脉粥样硬化等与内皮相关的疾病模型。原代培养的生物学行为依赖于供体，数据不易重现；内皮细胞系是新兴的选择，特别是人真皮微血管内皮细胞（HMEC-1），应经过验证后才能替代原代细胞用于研究 HDL 的功能。
方法
通过相差显微镜和 HMEC-1 的流式细胞术评估细胞的形态、大小和颗粒度。通过流式细胞术测定 TNF-α 刺激后粘附分子 ICAM-1 和 VCAM-1，以及内皮标志物 CD105 内格林和 HDL 受体 SR-BI。通过使用 Alexa Fluor 488 标记的 HDL 进行共聚焦显微镜观察，证明 HDL 蛋白的内化。将 HUVEC 用作参考，与 HMEC-1 比较其特征。
结果
HMEC-1 和 HUVEC 具有相似的形态、大小和颗粒度。HMEC-1 表达内皮标志物，如 HUVEC，以及功能性 SR-B1 受体，因为该细胞系能够内化 HDL 颗粒。HMEC-1 在 TNF-α 刺激后有效增加 ICAM-1 和 VCAM-1 的表达。HUVEC 对 TNF-α 刺激的敏感性更高，但 ICAM-1 和 VCAM-1 的表达范围不如 HMEC-1 均匀，这可能是由于前者的生物学变异性。最后，HDL 共孵育可减弱 HMEC-1 中粘附分子的表达。
结论
HMEC-1 具有与 HUVEC 相似的内皮细胞特征，是一种适合评估 HDL 对内皮功能的细胞系。
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