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

Alex L. Wilkinson; Megan E. Bannister; Ayla O'Keeffe; Chris J. Weston; Patricia F. Lalor; Shishir Shetty; Daniel A. Patten

doi:10.1016/j.xpro.2024.103431

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

Alex L. Wilkinson
, Megan E. Bannister
, Ayla O'Keeffe
, Chris J. Weston
, Patricia F. Lalor
, Shishir Shetty^*
, Daniel A. Patten^*

^*Corresponding author for this work

Immunology and Immunotherapy

Research output: Contribution to journal › Article › peer-review

140 Downloads (Pure)

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

Original language	English
Article number	103431
Number of pages	20
Journal	STAR Protocols
Volume	5
Issue number	4
Early online date	5 Nov 2024
DOIs	https://doi.org/10.1016/j.xpro.2024.103431
Publication status	Published - 20 Dec 2024

Bibliographical note

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.xpro.2024.103431Licence: Creative Commons: Attribution (CC BY)

WilkinsonA2024ProtocolFinal published version, 5.19 MBLicence: Creative Commons: Attribution (CC BY)

Endothelial scavenger receptors shape the immune microenvironment in hepatocellular carcinoma
Shetty, S. (Principal Investigator)
Cancer Research UK
1/09/20 → 31/08/26
Project: Research

Cite this

@article{d5a3b50363104205bc4bb3e019d0344b,

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

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",

author = "Wilkinson, \{Alex L.\} and Bannister, \{Megan E.\} and Ayla O'Keeffe and Weston, \{Chris J.\} and Lalor, \{Patricia F.\} and Shishir Shetty and Patten, \{Daniel A.\}",

note = "Copyright {\textcopyright} 2024 The Author(s). Published by Elsevier Inc.",

year = "2024",

month = dec,

day = "20",

doi = "10.1016/j.xpro.2024.103431",

language = "English",

volume = "5",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Elsevier",

number = "4",

}

TY - JOUR

T1 - Protocol to study monocyte transmigration across primary human liver endothelial cells under physiological shear flow conditions in vitro

AU - Wilkinson, Alex L.

AU - Bannister, Megan E.

AU - O'Keeffe, Ayla

AU - Weston, Chris J.

AU - Lalor, Patricia F.

AU - Shetty, Shishir

AU - Patten, Daniel A.

PY - 2024/12/20

Y1 - 2024/12/20

N2 - 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

AB - 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

U2 - 10.1016/j.xpro.2024.103431

DO - 10.1016/j.xpro.2024.103431

M3 - Article

C2 - 39504247

SN - 2666-1667

VL - 5

JO - STAR Protocols

JF - STAR Protocols

IS - 4

M1 - 103431

ER -

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

Abstract

Bibliographical note

UN SDGs

Access to Document

Fingerprint

Projects

Endothelial scavenger receptors shape the immune microenvironment in hepatocellular carcinoma

Cite this