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

Ségolène Ladaigue; Vincent Paget; Anne-Charlotte Lefranc; Monica Quitoco; Emilie Bacquer; Fabien Milliat; Olivier Guipaud

doi:10.1016/j.xpro.2023.102388

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

STAR Protoc. 2023 Sep 15;4(3):102388. doi: 10.1016/j.xpro.2023.102388. Epub 2023 Jun 27.

Authors

Ségolène Ladaigue¹, Vincent Paget², Anne-Charlotte Lefranc², Monica Quitoco², Emilie Bacquer², Fabien Milliat², Olivier Guipaud³

Affiliations

¹ Institut de Radioprotection et de Sûreté Nucléaire (IRSN), PSE-SANTE/SERAMED/LRMed, 92260 Fontenay-aux-Roses, France; Sorbonne University, Doctoral College, 75005 Paris, France.
² Institut de Radioprotection et de Sûreté Nucléaire (IRSN), PSE-SANTE/SERAMED/LRMed, 92260 Fontenay-aux-Roses, France.
³ Institut de Radioprotection et de Sûreté Nucléaire (IRSN), PSE-SANTE/SERAMED/LRMed, 92260 Fontenay-aux-Roses, France. Electronic address: olivier.guipaud@irsn.fr.

Abstract

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.

Keywords: Cell Culture; Cell-based Assays; Immunology; Microscopy.

MeSH terms

Cell Movement
Chemotaxis
Endothelial Cells
Humans
Monocytes*
Transendothelial and Transepithelial Migration*