Abstract

Artificial 3-dimensional (3D) cell culture systems, which mimic the extracellular matrix (ECM), hold great potential as models to study cellular processes under controlled conditions. The natural ECM is a 3D structure composed of a fibrous hydrogel that provides both mechanical and biochemical cues to instruct cell behavior. Here we present an ECM-mimicking genetically engineered protein-based hydrogel as a 3D cell culture system that combines several key features: (1) Mild and straightforward encapsulation meters (1) ease of ut I am not so sure.encapsulation of the cells, without the need of an external crosslinker. (2) Supramolecular assembly resulting in a fibrous architecture that recapitulates some of the unique mechanical characteristics of the ECM, i.e. strain-stiffening and self-healing behavior. (3) A modular approach allowing controlled incorporation of the biochemical cue density (integrin binding RGD domains). We tested the gels by encapsulating MG-63 osteoblastic cells and found that encapsulated cells not only respond to higher RGD density, but also to overall gel concentration. Cells in 1% and 2% (weight fraction) protein gels showed spreading and proliferation, provided a relative RGD density of at least 50%. In contrast, in 4% gels very little spreading and proliferation occurred, even for a relative RGD density of 100%. The independent control over both mechanical and biochemical cues obtained in this modular approach renders our hydrogels suitable to study cellular responses under highly defined conditions.

Highlights

  • In natural tissues, most cells interact with the native extracellular matrix (ECM) in a 3-dimensional (3D) environment [1,2]

  • We tested the gels by encapsulating MG-63 osteoblastic cells and found that encapsulated cells respond to higher RGD density, and to overall gel concentration

  • The independent control over both mechanical and biochemical cues obtained in this modular approach renders our hydrogels suitable to study cellular responses under highly defined conditions

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Summary

Introduction

Most cells interact with the native extracellular matrix (ECM) in a 3-dimensional (3D) environment [1,2]. The ECM, a fibrous mesh of high complexity and hierarchy, ensures proper molecular structure, functional bioactivity, and mechanical support for cells [2]. Mutual cell–ECM interactions form a dynamic regulatory system, directing cell behavior [1] and thereby influencing tissue formation and regeneration [3]. Current knowledge about cell–matrix interactions is mostly based on 2-dimensional (2D) in vitro studies. Culturing cells in a monolayer does not accurately represent the PLOS ONE | DOI:10.1371/journal.pone.0155625. Culturing cells in a monolayer does not accurately represent the PLOS ONE | DOI:10.1371/journal.pone.0155625 May 25, 2016

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