A microfluidic device for measuring cell migration towards substrate-bound and soluble chemokine gradients.
Cellular locomotion is a central hallmark of eukaryotic life. It is governed by cell-extrinsic molecular factors, which can either emerge in the soluble phase or as immobilized, often adhesive ligands. To encode for direction, every cue must be present as a spatial or temporal gradient. Here, we developed a microfluidic chamber that allows measurement of cell migration in combined response to surface immobilized and soluble molecular gradients. As a proof of principle we study the response of dendritic cells to their major guidance cues, chemokines. The majority of data on chemokine gradient sensing is based on in vitro studies employing soluble gradients. Despite evidence suggesting that in vivo chemokines are often immobilized to sugar residues, limited information is available how cells respond to immobilized chemokines. We tracked migration of dendritic cells towards immobilized gradients of the chemokine CCL21 and varying superimposed soluble gradients of CCL19. Differential migratory patterns illustrate the potential of our setup to quantitatively study the competitive response to both types of gradients. Beyond chemokines our approach is broadly applicable to alternative systems of chemo- and haptotaxis such as cells migrating along gradients of adhesion receptor ligands vs. any soluble cue.
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Jan
J
Schwarz
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Veronika
V
Bierbaum
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Jack
J
Merrin
Department of Biosystems Science and Engineering, ETH Zürich, 4058, Basel, Switzerland.
Tino
T
Frank
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Robert
R
Hauschild
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Tobias
T
Bollenbach
Department of Biosystems Science and Engineering, ETH Zürich, 4058, Basel, Switzerland.|Institute for Molecular Engineering, The University of Chicago, Chicago, IL, 60637, USA.
Savaş
S
Tay
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.
Michael
M
Sixt
Institute of Science and Technology Austria (IST Austria), 3400, Klosterneuburg, Austria.|Department of Biomedicine and Neurology Department, University Hospital Basel, 4031, Basel, Switzerland.
Matthias
M
Mehling
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D054415:Chemokine CCL19 / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D054421:Chemokine CCL21 / Q000737:chemistry / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects*; D003713:Dendritic Cells / Q000166:cytology / Q000502:physiology*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D056656:Lab-On-A-Chip Devices; D051379:Mice; D008810:Mice, Inbred C57BL; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence; D038761:Photobleaching; D013379:Substrate Specificity
D054415:Chemokine CCL19; D054421:Chemokine CCL21; D055767:Immobilized Proteins; D016650:Fluorescein-5-isothiocyanate