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2015 ; 7
(11
): 1454-65
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Fibronectin fibrillogenesis facilitates mechano-dependent cell spreading, force
generation, and nuclear size in human embryonic fibroblasts
#MMPMID26412391
Scott LE
; Mair DB
; Narang JD
; Feleke K
; Lemmon CA
Integr Biol (Camb)
2015[Nov]; 7
(11
): 1454-65
PMID26412391
show ga
Cells respond to mechanical cues from the substrate to which they are attached.
These mechanical cues drive cell migration, proliferation, differentiation, and
survival. Previous studies have highlighted three specific mechanisms through
which substrate stiffness directly alters cell function: increasing stiffness
drives (1) larger contractile forces; (2) increased cell spreading and size; and
(3) altered nuclear deformation. While studies have shown that substrate
mechanics are an important cue, the role of the extracellular matrix (ECM) has
largely been ignored. The ECM is a crucial component of the mechanosensing system
for two reasons: (1) many ECM fibrils are assembled by application of
cell-generated forces, and (2) ECM proteins have unique mechanical properties
that will undoubtedly alter the local stiffness sensed by a cell. We specifically
focused on the role of the ECM protein fibronectin (FN), which plays a critical
role in de novo tissue production. In this study, we first measured the effects
of substrate stiffness on human embryonic fibroblasts by plating cells onto
microfabricated pillar arrays (MPAs) of varying stiffness. Cells responded to
increasing substrate stiffness by generating larger forces, spreading to larger
sizes, and altering nuclear geometry. These cells also assembled FN fibrils
across all stiffnesses, with optimal assembly occurring at approximately 6 kPa.
We then inhibited FN assembly, which resulted in dramatic reductions in
contractile force generation, cell spreading, and nuclear geometry across all
stiffnesses. These findings suggest that FN fibrils play a critical role in
facilitating cellular responses to substrate stiffness.