Protocol abstract
Nature Protocols 1, - 1379 - 1391 (2006)
Published online: 2 November 2006 | doi:10.1038/nprot.2006.203
Subject Categories: Cell and tissue culture | Model organisms
Micropatterned cell cultures on elastic membranes as an in vitro model of myocardium
Patrizia Camelliti1,3, John O Gallagher2,3, Peter Kohl1 & Andrew D McCulloch2
Abstract
We describe here a new in vitro protocol for structuring cardiac cell cultures to mimic important aspects of the in vivo ventricular myocardial phenotype by controlling the location and mechanical environment of cultured cells. Microlithography is used to engineer microstructured silicon metal wafers. Those are used to fabricate either microgrooved silicone membranes or silicone molds for microfluidic application of extracellular matrix proteins onto elastic membranes (involving flow control at micrometer resolution). The physically or microfluidically structured membranes serve as a cell culture growth substrate that supports cell alignment and allows the application of stretch. The latter is achieved with a stretching device that can deliver isotropic or anisotropic stretch. Neonatal ventricular cardiomyocytes, grown on these micropatterned membranes, develop an in vivo–like morphology with regular sarcomeric patterns. The entire process from fabrication of the micropatterned silicon metal wafers to casting of silicone molds, microfluidic patterning and cell isolation and seeding takes approximately 7 days.
- Department of Physiology, Anatomy and Genetics, Parks Road, Oxford OX1 3PT, UK.
- Department of Bioengineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
- These authors contributed equally to this work.
Correspondence to: Peter Kohl1 e-mail: peter.kohl@physiol.ox.ac.uk
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