Substrate topography induces a crossover from 2D to 3D behavior in fibroblast migration.
In a three-dimensional environment, cells migrate through complex topographical features. Using microstructured substrates, we investigate the role of substrate topography in cell adhesion and migration. To do so, fibroblasts are plated on chemically identical substrates composed of microfabricated pillars. When the dimensions of the pillars (i.e., the diameter, length, and spacing) are varied, migrating cells encounter alternating flat and rough surfaces that depend on the spacing between the pillars. Consequently, we show that substrate topography affects cell shape and migration by modifying cell-to-substrate interactions. Cells on micropillar substrates exhibit more elongated and branched shapes with fewer actin stress fibers compared with cells on flat surfaces. By analyzing the migration paths in various environments, we observe different mechanisms of cell migration, including a persistent type of migration, that depend on the organization of the topographical features. These responses can be attributed to a spatial reorganization of the actin cytoskeleton due to physical constraints and a preferential formation of focal adhesions on the micropillars, with an increased lifetime compared to that observed on flat surfaces. By changing myosin II activity, we show that actomyosin contractility is essential in the cellular response to micron-scale topographic signals. Finally, the analysis of cell movements at the frontier between flat and micropillar substrates shows that cell transmigration through the micropillar substrates depends on the spacing between the pillars.
Ghibaudo
Marion
M
Laboratoire Matière et Systèmes Complexes (MSC), Université Paris Diderot and Centre National de la Recherche Scientifique, Unite Mixte de Recherche 7057, Paris, France.
Trichet
Léa
L
Le Digabel
Jimmy
J
Richert
Alain
A
Hersen
Pascal
P
Ladoux
Benoît
B
D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D000205:Actomyosin / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D024462:Myosin Type II / Q000378:metabolism; D054457:Tissue Scaffolds; D014162:Transfection; D014741:Video Recording; D016596:Vinculin / Q000378:metabolism
D000199:Actins; C485184:enhanced green fluorescent protein; D016596:Vinculin; D049452:Green Fluorescent Proteins; D000205:Actomyosin; D024462:Myosin Type II