naa a22 4500 528785044 20180924065518.0 cr unu---uuuuu 180924e20100323xx s 000 0 eng 10.3929/ethz-b-000017498 doi 10.1371/journal.pone.0009445 doi (ETHRESEARCH)oai:www.research-collection.ethz.ch:20.500.11850/17498 Dimensionality Controls Cytoskeleton Assembly and Metabolism of Fibroblast Cells in Response to Rigidity and Shape [Elektronische Daten] [Mirjam Ochsner, Marcus Textor, Viola Vogel, Michael L. Smith] PLoS ONE Open access ethresearch Background Various physical parameters, including substrate rigidity, size of adhesive islands and micro-and nano-topographies, have been shown to differentially regulate cell fate in two-dimensional (2-D) cell cultures. Cells anchored in a three-dimensional (3-D) microenvironment show significantly altered phenotypes, from altered cell adhesions, to cell migration and differentiation. Yet, no systematic analysis has been performed that studied how the integrated cellular responses to the physical characteristics of the environment are regulated by dimensionality (2-D versus 3-D). Methodology/Principal Findings Arrays of 5 or 10 µm deep microwells were fabricated in polydimethylsiloxane (PDMS). The actin cytoskeleton was compared for single primary fibroblasts adhering either to microfabricated adhesive islands (2-D) or trapped in microwells (3-D) of controlled size, shape, and wall rigidity. On rigid substrates (Young's Modulus = 1 MPa), cytoskeleton assembly within single fibroblast cells occurred in 3-D microwells of circular, rectangular, square, and triangular shapes with 2-D projected surface areas (microwell bottom surface area) and total surface areas of adhesion (microwell bottom plus wall surface area) that inhibited stress fiber assembly in 2-D. In contrast, cells did not assemble a detectable actin cytoskeleton in soft 3-D microwells (20 kPa), regardless of their shapes, but did so on flat, 2-D substrates. The dependency on environmental dimensionality was also reflected by cell viability and metabolism as probed by mitochondrial activities. Both were upregulated in 3-D cultured cells versus cells on 2-D patterns when surface area of adhesion and rigidity were held constant. Conclusion/Significance These data indicate that cell shape and rigidity are not orthogonal parameters directing cell fate. The sensory toolbox of cells integrates mechanical (rigidity) and topographical (shape and dimensionality) information differently when cell adhesions are confined to 2-D or occur in a 3-D space. Creative Commons Attribution 3.0 Unported http://creativecommons.org/licenses/by/3.0 ethresearch Ochsner Mirjam joint author Textor Marcus joint author Vogel Viola joint author Smith Michael L. joint author PLoS ONE Lawrence, KS, USA : Public Library of Science 5 (3), p. e9445 1932-6203 http://hdl.handle.net/20.500.11850/17498 text/html WWW-Backlink auf das Repository (Open access) 1 Journal Article ethresearch ETHRESEARCH 856 40 http://hdl.handle.net/20.500.11850/17498 text/html WWW-Backlink auf das Repository (Open access) ETHRESEARCH 700 1- Ochsner Mirjam joint author ETHRESEARCH 700 1- Textor Marcus joint author ETHRESEARCH 700 1- Vogel Viola joint author ETHRESEARCH 700 1- Smith Michael L. joint author ETHRESEARCH 773 0- PLoS ONE Lawrence, KS, USA : Public Library of Science 5 (3), p. e9445 1932-6203 BK010053 XK010053 XK010000 ETHRESEARCH ETHRESEARCH ETHRESEARCH Journal Article Open access