Diffusion of chemokines away from their site of production results in the passive formation of chemokine gradients. We have recently shown that chemokine gradients can also be formed in an active manner, namely by atypical chemokine receptors (ACKRs) that scavenge chemokines locally. Here, we describe an advanced method that allows the visualization of leukocyte migration in a three-dimensional environment along a chemokine gradient that is actively established by cells expressing an ACKR. Initially developed to visualize the migration of dendritic cells along gradients of CCL19 or CCL21 that were actively shaped by an ACKR4-expressing cell line, we expect that this chamber system can be exploited to study many other combinations of atypical and conventional chemokine receptor-expressing cells.
Keywords: Atypical chemokine receptors; Cell migration; Chemokine gradient; Live-cell imaging; Time-lapse imaging.
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