TY - JOUR
T1 - Substrate stiffness and matrix composition coordinately control the differentiation of liver progenitor cells
AU - Kourouklis, Andreas P.
AU - Kaylan, Kerim B.
AU - Underhill, Gregory H.
N1 - Funding Information:
We gratefully acknowledge Hélène Strick-Marchand and Mary C. Weiss (Institut Pasteur) for providing BMEL cells. We also acknowledge Roberto A. Eguiluz and Deborah E. Leckband (University of Illinois at Urbana-Champaign) for advice and assistance with traction force microscopy, Ning Wang (University of Illinois at Urbana-Champaign) for providing the original traction force analysis script, and Austin Cyphersmith and Mayandi Sivaguru (University of Illinois at Urbana-Champaign) for assistance with microscopy. This work was supported by start-up funding from the University of Illinois at Urbana-Champaign.
PY - 2016/8/1
Y1 - 2016/8/1
N2 - Recent approaches have utilized microfabricated platforms to examine combinations of microenvironmental signals that regulate stem and progenitor cell differentiation. However, the majority of these efforts have focused on the biochemical properties of extracellular matrix (ECM) or soluble factors without simultaneously exploring the biomechanical effects of cell-substrate interactions. To address this need, we combined a high-throughput approach for the analysis of combinatorial ECM cues with substrates of modular stiffness and traction force microscopy. This integrated approach enabled the characterization of cell-generated traction stress and phenotypic expression in response to ECM cues. We investigated the impact of substrate stiffness and ECM composition on the differentiation of bipotential mouse embryonic liver (BMEL) progenitor cells. We observed that hepatocyte differentiation was primarily regulated by ECM composition, and cholangiocyte differentiation was cooperatively influenced by ECM proteins and stiffness properties. In particular, stiffness-mediated cholangiocyte differentiation was observed for cells cultured on fibronectin, while collagen IV promoted differentiation independent of substrate stiffness. We demonstrated the influence of cell contractility and traction stress in early cholangiocyte specification and further uncovered the roles of ERK and ROCK in this differentiation process. Overall, these findings illustrate the involvement of biomechanical signals in liver progenitor differentiation. Further, this approach could enable investigations for a broad range of cell types and ECM proteins, providing an integrated platform for evaluating the combinatorial effects of biochemical and biophysical signals in cell differentiation.
AB - Recent approaches have utilized microfabricated platforms to examine combinations of microenvironmental signals that regulate stem and progenitor cell differentiation. However, the majority of these efforts have focused on the biochemical properties of extracellular matrix (ECM) or soluble factors without simultaneously exploring the biomechanical effects of cell-substrate interactions. To address this need, we combined a high-throughput approach for the analysis of combinatorial ECM cues with substrates of modular stiffness and traction force microscopy. This integrated approach enabled the characterization of cell-generated traction stress and phenotypic expression in response to ECM cues. We investigated the impact of substrate stiffness and ECM composition on the differentiation of bipotential mouse embryonic liver (BMEL) progenitor cells. We observed that hepatocyte differentiation was primarily regulated by ECM composition, and cholangiocyte differentiation was cooperatively influenced by ECM proteins and stiffness properties. In particular, stiffness-mediated cholangiocyte differentiation was observed for cells cultured on fibronectin, while collagen IV promoted differentiation independent of substrate stiffness. We demonstrated the influence of cell contractility and traction stress in early cholangiocyte specification and further uncovered the roles of ERK and ROCK in this differentiation process. Overall, these findings illustrate the involvement of biomechanical signals in liver progenitor differentiation. Further, this approach could enable investigations for a broad range of cell types and ECM proteins, providing an integrated platform for evaluating the combinatorial effects of biochemical and biophysical signals in cell differentiation.
KW - Cellular microarrays
KW - Extracellular matrix
KW - Liver progenitor cells
KW - Substrate stiffness
KW - Traction force microscopy
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U2 - 10.1016/j.biomaterials.2016.05.016
DO - 10.1016/j.biomaterials.2016.05.016
M3 - Article
C2 - 27235994
AN - SCOPUS:84969802809
SN - 0142-9612
VL - 99
SP - 82
EP - 94
JO - Biomaterials
JF - Biomaterials
ER -