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Transendothelial Migration

Endothelial cells (EC) line the lumen of blood and lymphatic vessels. Here these cells form a barrier and control tissue entry of cells and substances. The control over the entry of immune cells is mediated by a process termed transendothelial migration (TEM). Upon local inflammation endothelial cells attract and activate passing immune cells using chemokines and adhesion molecules. This activates immune cells and induces TEM, to alleviate inflammation. The TEM process in for example the invasion of tumor tissue by immune cells stands central to modern therapies like adoptive transfer of therapeutic T-cells or natural killer (NK) cells.

HUVEC, cultured under flow, F-actin (green), β-tubulin (orange), Nuclei (blue)
Automated tracking of segment.ai identified nuclei of PBMC on inflamed endothelial cells under flow
Transendothelial migration graph distinguishing surface migration and fully transmigrated immune cells.
Examples of surface migration and transmigration

Challenges

Transmigration experiments require dedicated expertise as well as systems for blood cell isolation and perfusion. In addition, identification of transmigrated cells thus far mostly relies on manual analysis, making TEM assays very demanding. The rapid and complex behavior of immune cells, especially when interacting with activated endothelium, leads to various outcomes such as cell polarization, surface migration, or transmigration, occurring over different timeframes. This complexity has historically made automated analysis highly challenging and time-consuming.

Solutions

One of NBIL’s specializations are immune cell and endothelial biology applications. We can provide blood cell isolations (Neutrophils, CD8+ T cells, NK cells), immune cell expansion and culture (e.g. CD8 T cells) from fresh donor blood, as well as endothelial cell culture. We also offer various options for TEM experiments ranging from perfused channels to complex micro physiological systems (MPS). Using Nikon AI assisted segmentation routines and automated tracking algorithms allows for individual immune cell tracking and automatic identification of transmigrated cells. Automating these, previously manual, analysis workflows considerably speeds up TEM assays and allows for higher throughput and increased cost effectiveness.

The NBIL is staffed with expert assay scientists & microscopists who will work with you to develop and execute customized image acquisition and analysis workflows to advance your research.

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