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

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 f...

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Autores principales: Ladaigue, Ségolène, Paget, Vincent, Lefranc, Anne-Charlotte, Quitoco, Monica, Bacquer, Emilie, Milliat, Fabien, Guipaud, Olivier
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331580/
https://www.ncbi.nlm.nih.gov/pubmed/37379221
http://dx.doi.org/10.1016/j.xpro.2023.102388
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author Ladaigue, Ségolène
Paget, Vincent
Lefranc, Anne-Charlotte
Quitoco, Monica
Bacquer, Emilie
Milliat, Fabien
Guipaud, Olivier
author_facet Ladaigue, Ségolène
Paget, Vincent
Lefranc, Anne-Charlotte
Quitoco, Monica
Bacquer, Emilie
Milliat, Fabien
Guipaud, Olivier
author_sort Ladaigue, Ségolène
collection PubMed
description 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)
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spelling pubmed-103315802023-07-11 Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system Ladaigue, Ségolène Paget, Vincent Lefranc, Anne-Charlotte Quitoco, Monica Bacquer, Emilie Milliat, Fabien Guipaud, Olivier STAR Protoc Protocol 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) Elsevier 2023-06-27 /pmc/articles/PMC10331580/ /pubmed/37379221 http://dx.doi.org/10.1016/j.xpro.2023.102388 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ladaigue, Ségolène
Paget, Vincent
Lefranc, Anne-Charlotte
Quitoco, Monica
Bacquer, Emilie
Milliat, Fabien
Guipaud, Olivier
Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title_full Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title_fullStr Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title_full_unstemmed Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title_short Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
title_sort protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10331580/
https://www.ncbi.nlm.nih.gov/pubmed/37379221
http://dx.doi.org/10.1016/j.xpro.2023.102388
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