Transendothelial migration induces differential migration dynamics of leukocytes in tissue matrix

Abstract

Leukocyte extravasation into inflamed tissue is a complex process that is difficult to capture as a whole in vitro. We employed a blood-vessel-on-a-chip model in which endothelial cells were cultured in a tube-like lumen in a collagen-1 matrix. The vessels are leak-tight, creating a barrier for molecules and leukocytes. Addition of inflammatory cytokine TNF-α caused vasoconstriction, actin remodelling and upregulation of ICAM-1. Introducing leukocytes into the vessels allowed real-time visualisation of leukocyte migration across the vessel wall, into the extracellular matrix. Individual cell tracking over time distinguished striking differences in migratory behaviour between T-cells and neutrophils. Neutrophils cross the endothelial layer more efficiently than T-cells, but upon entering the matrix, neutrophils display high speed but low persistence, whereas T-cells migrate with low speed and rather linear migration. In conclusion, 3D imaging in real-time of leukocyte extravasation in a vessel-on-a-chip enables detailed qualitative and quantitative analysis of different stages of the full leukocyte extravasation process in a single assay.

Endothelial transmigration hotspots limit vascular leakage through heterogeneous expression of ICAM1

“…No change in the degree of heterogeneity of any of the adhesion molecules measured was found (Figure S2D). Using a vessel-on-achip model, developed by our lab 40 , we confirmed ICAM-1 heterogeneous distribution in a 3D inflamed vessel (Figure 3D and S2E). Clinically obtained samples of chronically inflamed human mesentery of inflammatory bowel disease patients showed ICAM-1 cell-to-cell heterogeneity in small veins (Figure 3E).…”

“…Vessel-on-a-chip samples were cultured and imaged as described here 40 . Patient tissue samples were received from the department of Pathology of the Amsterdam UMC, location AMC.…”

Development of a biofabricated 3D in vitro vessel model for investigating transendothelial migration in stem cell therapy

“…Bogseth et al provided a review of similar in vitro models focusing on vascular inflammation, transendothelial migration, and extravasation of cells [19]. Previously developed models are dominated by conventional transwell assays or microfluidic chips, which are mainly used for the investigation of leukocyte transendothelial migration [14,19,75]. An advantage of microfluidic chips lies within the facilitated microscopy and cell tracking [14,76,77].…”

“…In general, transendothelial migration of cells occurs in capillaries or post-capillary venules, as the laminar flow velocity of the blood and cells is sufficiently low to allow the rolling and catching of cells by selectins and integrins [12,13]. The rolling and adhesion of cells on the endothelial layer takes only seconds, while the transendothelial migration itself and the crossing of the basal lamina lasts 2-3 and 15-20 mins, respectively [13,14].…”

“…Furthermore, they should enable the biochemical (cytokines, hormones), physiochemical (oxygen/temperature level) or mechanical (stretch, shear stress) stimulation of the vessel model, e.g. to mimic inflammatory responses in vitro and facilitate the in situ imaging [14,27,28].…”