Mechanical regulation of cell function with

geometrically modulated elastomeric

substrates

SUMMARY
Micromolded elastomeric micropost arrays
modulate substrate rigidity.
Micropost rigidity affect cell morphology, focal
adhesions (FA), cytoskeletal contractility and
stem cell differentiation.

Manufacturing micropost

INTRODUCTION
Cell function is regulated primarily by extracellular
stimuli.
Recent advances suggest that the rigidity of the
extracellular matrix (ECM) can mediate cell signaling,
proliferation, differentiation and migration.
Culturing cells on hydrogels based on ECM proteins
has a strong impact on cell adhesion, morphology and
function.
Substrate rigidity can modulate many cellular functons
including stem cell differentiation.

Results
Micromolded elastomeric micropost arrays
can decouple substrate rigidity from adhesive
and surface properties.
Post height determines the bending degree in
response to horizontal traction force. Rigidity
characterized by computing the nominal
spring constant K (1.31-1.556 nN/m).

a) FEM analysis of bending in response to a horizontal traction force (20

nN).
b) Micropost deflection as function of traction force.
c) Nominal spring constant as function of post height L (FEM analysis
(bars) and Euler-Bernouilli beam theory (curve)).

 Different behaviour in hMSCs cultured on

high/low rigidity microposts:
high rigidity leads to well spread cells, with
prominent, highly organized actin stress fibers and
large FA.
Low rigidity leads to a rounded morphology with
prominent microvilli.
high rigidity:
L = 0,97 m,
K = 1,556 nN/m;
medium rigiditiy:
L = 6.1 m,
K = 18.16 nN/m
low rigidity:
L = 12.9 m,
K = 1.90 nN /m

 Strong correlations between:

FA and cell spreading regardless of micropost
rigidity, and
Traction force and cell spreading.

Advantages of using micropost over

hydrogels
Responses similar to those reported on hydrogels
of varying rigidities, but with rigidity sensing at a
micrometer scale (between focal adhesions).
Measured subcellular traction forces could be
attributed to FA. This allows us to map traction
forces in individual FA and spatially quantify
subcellular distributios of FA area, traction force
and FA stress.

Traction force as a function of FA area for different

rigidities; hMSCs (top) and hUVECs (bottom)

FA stress anisotropy for hMSCs and HUVECs.

Average FA stress per cell increased with micropost
rigidity, but different for the 2 types.

FA stress computed as the slope of

the linear fit in traction force vs FA
area plots.

There may be multiple ways for

cells to mechanically adapt to
their environment.