title: Microfabrication of a platform to measure and manipulate the mechanics of engineered microtissues.
abstract: Engineered tissues can be used to understand fundamental features of biology, develop organotypic in vitro model systems, and as engineered tissue constructs for replacing damaged tissue in vivo. However, a key limitation is an inability to test the wide range of parameters that might impact the engineered tissue in a high-throughput manner and in an environment that mimics the three-dimensional (3D) native architecture. We developed a microfabricated platform to generate arrays of microtissues embedded within 3D micropatterned matrices. Microcantilevers simultaneously constrain microtissue formation and report forces generated by the microtissues in real time, opening the possibility to use high-throughput, low-volume screening for studies on engineered tissues. Thanks to the micrometer scale of the microtissues, this platform is also suitable for high-throughput monitoring of drug-induced effect on architecture and contractility in engineered tissues. Moreover, independent variations of the mechanical stiffness of the cantilevers and collagen matrix allow the measurement and manipulation of the mechanics of the microtissues. Thus, our approach will likely provide valuable opportunities to elucidate how biomechanical, electrical, biochemical, and genetic/epigenetic cues modulate the formation and maturation of 3D engineered tissues. In this chapter, we describe the microfabrication, preparation, and experimental use of such microfabricated tissue gauges.
authors.affiliation: Laboratory of Materials and Physical Engineering, CNRS UMR5628, and Grenoble Institute of Technology, Grenoble, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Ramade
authors.affiliation: HHMI Janelia Farm Research Campus, Ashburn, Virginia, USA.
authors.forename: Wesley R
authors.initials: WR
authors.lastname: Legant
authors.affiliation: Laboratory of Materials and Physical Engineering, CNRS UMR5628, and Grenoble Institute of Technology, Grenoble, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Picart
authors.affiliation: Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Chen
authors.affiliation: Laboratory of Materials and Physical Engineering, CNRS UMR5628, and Grenoble Institute of Technology, Grenoble, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Boudou
mesh_terms: D016475:3T3 Cells / Q000166:cytology; D000818:Animals; D001696:Biomechanical Phenomena / Q000502:physiology*; D018929:Cell Culture Techniques; D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D055119:Elastic Modulus; D004558:Electric Stimulation; D004852:Epoxy Compounds / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D051379:Mice; D055616:Microtechnology; D018482:Muscle, Skeletal / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology; D011108:Polymers / Q000737:chemistry; D051381:Rats; D013314:Stress, Mechanical; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes; D004852:Epoxy Compounds; D011108:Polymers; C520895:SU-8 compound; C013830:baysilon
keywords: Biomechanics; Cell mechanics; Collagen; PDMS; Tissue engineering