Guidelines for evaluating endothelial function in vascular tissue

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McCarthy, Cameron G. and Aalkjær, Christian and Bagher, Pooneh and Beyer, Andreas M. and Boedtkjer, Ebbe and Bomfim, Gisele F. and Breslin, Jerome W. and Briones, Ana M. and Castorena-Gonzalez, Jorge A. and Costa, Tiago J. and Dai, Zhiyu and Davel, Ana P. and Earley, Scott and Freed, Julie K. and Garland, Christopher and Isakson, Brant E. and Jepps, Thomas A. and Kalucka, Joanna and Lavanderos, Boris and Makino, Ayako and Norton, Charles E. and Segal, Steven S. and Tan, Wenbin and Trask, Aaron J. and Wilson, Calum and Zawieja, Scott D. and Wenceslau, Camilla F. (2026) Guidelines for evaluating endothelial function in vascular tissue. American Journal of Physiology - Heart and Circulatory Physiology, 330 (5). H1600–H1672. ISSN 0363-6135 (https://doi.org/10.1152/ajpheart.00656.2025)

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Abstract

The endothelium plays a central role in maintaining vascular homeostasis by orchestrating vascular tone, inflammation, healing, permeability, and thrombosis. Assessing endothelial function in vascular tissue is essential for understanding the cellular and molecular mechanisms underlying cardiovascular physiology and pathology. Traditional approaches, such as wire and pressure myography, have been instrumental in defining endothelium-dependent responses and identifying key pharmacological targets. However, the complexity and heterogeneity of endothelial cells across vascular beds, and their dynamic phenotypic changes in health and disease, necessitate the incorporation of new investigative strategies. Emerging methodologies, including bulk and single-cell transcriptomics, proteomics, and advanced imaging, now provide unprecedented insights into endothelial cell diversity and function. A team of leading experts in the field, who collectively reached a consensus on the most widely used techniques to evaluate endothelial function, developed these guidelines. The document establishes best practices for assessing endothelial function, from endothelial cell cultures to isolated vascular tissues, integrating conventional functional assays with modern molecular approaches. By fostering methodological consistency and embracing innovation, our goal is to enhance rigor, reproducibility, understanding, and discovery in endothelial biology.

ORCID iDs

McCarthy, Cameron G., Aalkjær, Christian, Bagher, Pooneh, Beyer, Andreas M., Boedtkjer, Ebbe, Bomfim, Gisele F., Breslin, Jerome W., Briones, Ana M., Castorena-Gonzalez, Jorge A., Costa, Tiago J., Dai, Zhiyu, Davel, Ana P., Earley, Scott, Freed, Julie K., Garland, Christopher, Isakson, Brant E., Jepps, Thomas A., Kalucka, Joanna, Lavanderos, Boris, Makino, Ayako, Norton, Charles E., Segal, Steven S., Tan, Wenbin, Trask, Aaron J., Wilson, Calum ORCID logoORCID: https://orcid.org/0000-0003-2500-0632, Zawieja, Scott D. and Wenceslau, Camilla F.;
CORE (COnnecting REpositories)