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Protocol to study monocyte transmigration across primary human liver endothelial cells under physiological shear flow conditions in vitro

Year: 2024

Type: article

Abstract: Modeling immune cell recruitment by liver endothelial cells in vitro is important to better understand the pathology of chronic inflammatory liver diseases and cancers. Here, we present a protocol for the study of monocyte transmigration across activated primary human liver endothelial cells, under physiological flow conditions. We describe primary endothelial cell isolation from human liver tissues and monocyte isolation from human blood. We then detail the shear flow-based assay and subsequent analysis of the different stages of monocyte transmigration. For complete details on the use and execution of this protocol, please refer to Wilkinson et al. 1 • Step-by-step guide for isolating and culturing human liver endothelial cells • Protocol for isolating monocytes from human blood • Undertaking shear flow-based assays of monocytes across liver endothelial cells • Quantitative analysis of monocyte transmigration route Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Modeling immune cell recruitment by liver endothelial cells in vitro is important to better understand the pathology of chronic inflammatory liver diseases and cancers. Here, we present a protocol for the study of monocyte transmigration across activated primary human liver endothelial cells, under physiological flow conditions. We describe primary endothelial cell isolation from human liver tissues and monocyte isolation from human blood. We then detail the shear flow-based assay and subsequent analysis of the different stages of monocyte transmigration.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103431