Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system

Autor: Ségolène Ladaigue, Vincent Paget, Anne-Charlotte Lefranc, Monica Quitoco, Emilie Bacquer, Fabien Milliat, Olivier Guipaud
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 3, Pp 102388- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2023.102388
Popis: Summary: In vitro modeling of the different steps of immune cell recruitment is essential to decipher the role of endothelial cells in this process. Here, we present a protocol for the assessment of human monocyte transendothelial migration using a live cell imaging system. We describe steps for culture of fluorescent monocytic THP-1 cells and chemotaxis plate preparation with HUVEC monolayers. We then detail real-time analysis using the IncuCyte® S3 live-cell imaging system, image analysis, and assessment of transendothelial migration rates.For complete details on the use and execution of this protocol, please refer to Ladaigue et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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