Protocol to study monocyte transmigration across primary human liver endothelial cells under physiological shear flow conditions in vitro
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.<sup>1</sup>.
Authors
- Ayla O'Keeffe
- Chris J. Weston (ORCID: https://orcid.org/0000-0002-9651-1264)
- Daniel A. Patten (ORCID: https://orcid.org/0000-0002-8745-0578)
- Alex L. Wilkinson (ORCID: https://orcid.org/0000-0002-9506-8555)
- Shishir Shetty (ORCID: https://orcid.org/0000-0002-4729-2173)
- Patricia F. Lalor (ORCID: https://orcid.org/0000-0001-8733-5273)
- Megan E. Bannister
Institutions
- NIHR Birmingham Liver Biomedical Research Unit (GB)
- NIHR Birmingham Biomedical Research Centre (GB)
- University of Birmingham (GB)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-11-05
- DOI
- https://doi.org/10.1016/j.xpro.2024.103431
- Primary Topic
- Phagocytosis and Immune Regulation
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Wellcome Trust
- Cancer Research UK
- National Institute for Health and Care Research
- Department of Health and Social Care
- University of Birmingham
- Engineering and Physical Sciences Research Council
- NIHR Imperial Biomedical Research Centre
- Birmingham Biomedical Research Centre