A Microfluidic 3D Endothelium-on-a-Chip Model to Study Transendothelial Migration of T Cells in Health and Disease.

Luuk De Haan,Johnny Suijker,Lenie J Van Den Broek,Wouter Strijker,Ruthger Van Roey,Thomas Olivier,Karla Queiroz,Oliver Poeschke,Elissaveta Petrova,Nina Berges,Sakshi Garg

doi:10.3390/ijms22158234

Luuk De Haan, Johnny Suijker + Show 9 more

Open Access

https://doi.org/10.3390/ijms22158234

Copy DOI

Abstract

The recruitment of T cells is a crucial component in the inflammatory cascade of the body. The process involves the transport of T cells through the vascular system and their stable arrest to vessel walls at the site of inflammation, followed by extravasation and subsequent infiltration into tissue. Here, we describe an assay to study 3D T cell dynamics under flow in real time using a high-throughput, artificial membrane-free microfluidic platform that allows unimpeded extravasation of T cells. We show that primary human T cells adhere to endothelial vessel walls upon perfusion of microvessels and can be stimulated to undergo transendothelial migration (TEM) by TNFα-mediated vascular inflammation and the presence of CXCL12 gradients or ECM-embedded melanoma cells. Notably, migratory behavior was found to differ depending on T cell activation states. The assay is unique in its comprehensiveness for modelling T cell trafficking, arrest, extravasation and migration, all in one system, combined with its throughput, quality of imaging and ease of use. We envision routine use of this assay to study immunological processes and expect it to spur research in the fields of immunological disorders, immuno-oncology and the development of novel immunotherapeutics.

Highlights

The immune system is a complex system that provides protection against potential threats to the integrity of the human body [1]
An endothelial vessel that is stable over the course of the experiment is a crucial prerequisite for an effective T cell extravasation assay
We describe a novel 3D T cell extravasation and migration assay using a high-throughput microfluidic platform

Summary

Introduction

The immune system is a complex system that provides protection against potential threats to the integrity of the human body [1]. It consists of various cell types with immensely diverse and highly specific receptors to recognize these threats and distinguish self from non-self. One important step in the immunological cascade in both health and disease is the recruitment of T cells towards the site of inflammation [4,5] These T cells traverse the body through blood vessels of the circulatory system and are recruited by different factors.

Methods

Results

Conclusion