Generation of stable advective-diffusive chemokine gradients in a three-dimensional hydrogel

Bonneuil, W., Watson, D., Frattolin, J., Russell, M. J., Fasanella Masci, F., Bandara, M. , Brook, B. S., Nibbs, R. J.B. and Moore, J. (2022) Generation of stable advective-diffusive chemokine gradients in a three-dimensional hydrogel. AIP Advances, 12(2), 025121. (doi: 10.1063/5.0064947)

Abstract

Physiologic chemoattractant gradients are shaped by diffusion, advection, binding to an extracellular matrix, and removal by cells. Previous in vitro tools for studying these gradients and the cellular migratory response have required cells to be constrained to a 2D substrate or embedded in a gel devoid of fluid flow. Cell migration in fluid flow has been quantified in the absence of chemoattractant gradients and shown to be responsive to them, but there is a need for tools to investigate the synergistic, or antagonistic, effects of gradients and flow. We present a microfluidic chip in which we generated precisely controlled gradients of the chemokine CCL19 under advective-diffusive conditions. Using torque-actuated membranes situated between a gel region and the chip outlet, the resistance of fluid channels adjacent to the gel region could be modified, creating a controllable pressure difference across the gel at a resolution inferior to 10 Pa. Constant supply and removal of chemokine on either side of the chip facilitated the formation of stable gradients at Péclet numbers between −10 and +10 in a collagen type I hydrogel. The resulting interstitial flow was steady within 0.05 μm s−1 for at least 8 h and varied by less than 0.05 μm s−1 along the gel region. This method advances the physiologic relevance of the study of the formation and maintenance of molecular gradients and cell migration, which will improve the understanding of in vivo observations.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Nibbs, Professor Rob and Jayaweera Bandara, Dr Mikaila and Fasanella Masci, Dr Francesca
• Authors: Bonneuil, W., Watson, D., Frattolin, J., Russell, M. J., Fasanella Masci, F., Bandara, M., Brook, B. S., Nibbs, R. J.B., and Moore, J.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Life Sciences
• Research Centre: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Immunobiology
• Journal Name: AIP Advances
• Publisher: American Institute of Physics
• ISSN: 2158-3226
• ISSN (Online): 2158-3226
• Published Online: 16 February 2022
• Copyright Holders: Copyright © 2022 The Authors
• First Published: First published in AIP Advances 12(2): 025121
• Publisher Policy: Reproduced under a Creative Commons License

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
301499		Integrative Transport Phenomena in Chemokine Gradient Establishment	Robert Nibbs	Wellcome Trust (WELLCOTR)	206284/Z/17/Z	III - Immunology