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.
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Acknowledgements
We thank the following collaborators who contributed to the development of these protocols: S. Bhatia (Massachusetts Institute of Technology, Boston, Massachusetts), C. Flaim (University of California, San Diego, La Jolla, California), S. Gopalan and T. Borg (University of South Carolina, Columbia, South Carolina). Supported by the National Heart Lung and Blood Institute (R21 HL072160 and HL46345 to A.D.M.) and the UK Biotechnology and Biological Sciences Research Council (18561 to P.K.); P.C. is a Junior Research Fellow at Christ Church, Oxford; and P.K. is a British Heart Foundation Research Fellow.
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Camelliti, P., Gallagher, J., Kohl, P. et al. Micropatterned cell cultures on elastic membranes as an in vitro model of myocardium. Nat Protoc 1, 1379–1391 (2006). https://doi.org/10.1038/nprot.2006.203
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DOI: https://doi.org/10.1038/nprot.2006.203
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