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体外生理剪切流条件下研究单核细胞跨原代人肝内皮细胞迁移的实验方案。

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

作者信息

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

机构信息

Centre for Liver and Gastrointestinal Research, Institute of Immunology and Immunotherapy, University of Birmingham, B15 2TT Birmingham, UK.

Centre for Liver and Gastrointestinal Research, Institute of Immunology and Immunotherapy, University of Birmingham, B15 2TT Birmingham, UK; National Institute for Health Research (NIHR), Birmingham Biomedical Research Centre, Birmingham, UK.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103431. doi: 10.1016/j.xpro.2024.103431. Epub 2024 Nov 5.

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..

摘要

体外模拟肝内皮细胞募集免疫细胞对于更好地理解慢性炎症性肝病和癌症的病理过程非常重要。在此,我们提出了一种在生理流动条件下研究单核细胞跨活化原代人肝内皮细胞迁移的方案。我们描述了从人肝组织中分离原代内皮细胞以及从人血液中分离单核细胞的方法。然后,我们详细介绍了基于剪切流的检测方法以及对单核细胞迁移不同阶段的后续分析。有关本方案使用和执行的完整详细信息,请参考威尔金森等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a7fa/11577181/fbdc8ff3ac91/fx1.jpg

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