Assaying Macrophage Chemotaxis Using Fluid‐Walled Microfluidics. Issue 9 (19th July 2022)

• Record Type: Journal Article
• Authors: Deroy, Cyril; Rumianek, Agata N.; Wheeler, James H. R.; Nebuloni, Federico; Cook, Peter R.; Greaves, David R.; Walsh, Edmond J.
• Is Part Of: Advanced materials technologies. Volume 7:Issue 9(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-07-19
• Subjects: cell patterning -- fluid walls -- macrophage chemotaxis -- microfluidics; Materials science -- Periodicals; Technological innovations -- Periodicals; Materials science; Technological innovations; Periodicals; 620.1105
• DOI: 10.1002/admt.202200279
• Languages: English
• ISSNs: 2365-709X

Abstract

Abstract: While many tools exist to study immune‐cell chemotaxis in vitro, current methods often lack desirable features. Using fluid‐walled microfluidics, circuits are built around primary murine macrophages deposited in pre‐defined patterns on Petri dishes or microplates. Concentration gradients of complement component 5a (C5a) are established in flow‐free or flowing environments, cell migration imaged, and cell directionality and velocity correlated to calculated local C5a concentrations. In flow‐free circuits built around patterned macrophages, only cells nearest the C5a source migrate regardless of local attractant concentration. Conversely, in flowing circuits free from intercellular signaling and attractant degradation, only cells distant from the source migrate. In both systems, cells respond to lower C5a concentrations than previously reported (≈0.1 pM). Finally, macrophages follow instantly‐shifted gradients better than slowly‐shifting ones, suggesting that migration depends on both spatial and temporal responses to concentration. Abstract : A method to pattern cells freely on standard Petri dishes is introduced and microfluidic circuits are built around them. Concentration gradients of chemoattractant are then generated in both passive (flow‐free) and active (with flow) circuits, and cell behavior tracked and analyzed. The method is used to investigate murine bone marrow‐derived macrophage chemotaxis towards an inflammatory peptide (C5a), revealing interesting population dynamics.