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Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration

Physical features of the extracellular matrix (ECM) heavily influence cell migration strategies and efficiency. Migration in and on fibrous ECMs is of significant physiologic importance, but limitations in the ability to experimentally define the diameter, density, and alignment of native ECMs in vi...

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Autores principales: Wang, William Y., Pearson, Alexander T., Kutys, Matthew L., Choi, Colin K., Wozniak, Michele A., Baker, Brendon M., Chen, Christopher S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6481732/
https://www.ncbi.nlm.nih.gov/pubmed/31069329
http://dx.doi.org/10.1063/1.5052239
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author Wang, William Y.
Pearson, Alexander T.
Kutys, Matthew L.
Choi, Colin K.
Wozniak, Michele A.
Baker, Brendon M.
Chen, Christopher S.
author_facet Wang, William Y.
Pearson, Alexander T.
Kutys, Matthew L.
Choi, Colin K.
Wozniak, Michele A.
Baker, Brendon M.
Chen, Christopher S.
author_sort Wang, William Y.
collection PubMed
description Physical features of the extracellular matrix (ECM) heavily influence cell migration strategies and efficiency. Migration in and on fibrous ECMs is of significant physiologic importance, but limitations in the ability to experimentally define the diameter, density, and alignment of native ECMs in vitro have hampered our understanding of how these properties affect this basic cell function. Here, we designed a high-throughput in vitro platform that models fibrous ECM as collections of lines of cell-adhesive fibronectin on a flat surface to eliminate effects of dimensionality and topography. Using a microcontact printing approach to orthogonally vary line alignment, density, and size, we determined each factor's individual influence on NIH3T3 fibroblast migration. High content imaging and statistical analyses revealed that ECM alignment is the most critical parameter in influencing cell morphology, polarization, and migratory behavior. Specifically, increasing ECM alignment led cells to adopt an elongated uniaxial morphology and migrate with enhanced speed and persistence. Intriguingly, migration speeds were tightly correlated with the organization of focal adhesions, where cells with the most aligned adhesions migrated fastest. Highly organized focal adhesions and associated actin stress fibers appeared to define the number and location of protrusive fronts, suggesting that ECM alignment influences active Rac1 localization. Utilizing a novel microcontact-printing approach that lacks confounding influences of substrate dimensionality, mechanics, or differences in the adhesive area, this work highlights the effect of ECM alignment on orchestrating the cytoskeletal machinery that governs directed uniaxial cell migration.
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spelling pubmed-64817322019-05-08 Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration Wang, William Y. Pearson, Alexander T. Kutys, Matthew L. Choi, Colin K. Wozniak, Michele A. Baker, Brendon M. Chen, Christopher S. APL Bioeng Articles Physical features of the extracellular matrix (ECM) heavily influence cell migration strategies and efficiency. Migration in and on fibrous ECMs is of significant physiologic importance, but limitations in the ability to experimentally define the diameter, density, and alignment of native ECMs in vitro have hampered our understanding of how these properties affect this basic cell function. Here, we designed a high-throughput in vitro platform that models fibrous ECM as collections of lines of cell-adhesive fibronectin on a flat surface to eliminate effects of dimensionality and topography. Using a microcontact printing approach to orthogonally vary line alignment, density, and size, we determined each factor's individual influence on NIH3T3 fibroblast migration. High content imaging and statistical analyses revealed that ECM alignment is the most critical parameter in influencing cell morphology, polarization, and migratory behavior. Specifically, increasing ECM alignment led cells to adopt an elongated uniaxial morphology and migrate with enhanced speed and persistence. Intriguingly, migration speeds were tightly correlated with the organization of focal adhesions, where cells with the most aligned adhesions migrated fastest. Highly organized focal adhesions and associated actin stress fibers appeared to define the number and location of protrusive fronts, suggesting that ECM alignment influences active Rac1 localization. Utilizing a novel microcontact-printing approach that lacks confounding influences of substrate dimensionality, mechanics, or differences in the adhesive area, this work highlights the effect of ECM alignment on orchestrating the cytoskeletal machinery that governs directed uniaxial cell migration. AIP Publishing LLC 2018-12-26 /pmc/articles/PMC6481732/ /pubmed/31069329 http://dx.doi.org/10.1063/1.5052239 Text en © 2018 Author(s). 2473-2877/2018/2(4)/046107/16 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Articles
Wang, William Y.
Pearson, Alexander T.
Kutys, Matthew L.
Choi, Colin K.
Wozniak, Michele A.
Baker, Brendon M.
Chen, Christopher S.
Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title_full Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title_fullStr Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title_full_unstemmed Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title_short Extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
title_sort extracellular matrix alignment dictates the organization of focal adhesions and directs uniaxial cell migration
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6481732/
https://www.ncbi.nlm.nih.gov/pubmed/31069329
http://dx.doi.org/10.1063/1.5052239
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