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Low-temperature biopreservation and 3D tissue engineering present two differing routes towards eventual on-demand access to transplantable biologics, but recent advances in both fields present critical new opportunities for crossover between them. In this work, we demonstrate sub-zero centigrade preservation and revival of autonomously beating three-dimensional human induced pluripotent stem cell (hiPSC)-derived cardiac microtissues via isochoric supercooling, without the use of chemical cryoprotectants. We show that these tissues can cease autonomous beating during preservation and resume it after warming, that the supercooling process does not affect sarcomere structural integrity, and that the tissues maintain responsiveness to drug exposure following revival. Our work suggests both that functional three dimensional (3D) engineered tissues may provide an excellent high-content, low-risk testbed to study complex tissue biopreservation in a genetically human context, and that isochoric supercooling may provide a robust method for preserving and reviving engineered tissues themselves.
Bibliographical noteFunding Information:
Funding is gratefully acknowledged from the NSF Engineering Research Center for Advanced Technologies for Preservation of Biological Systems (ATP-Bio) NSF EEC #1941543. We would also like to thank Dr. Henrik Finsberg and Dr. Sam Wall (Simula Research Laboratory, Oslo, Norway) for the Python scripts used to extract beat information from calcium traces, Dr. Bruce Conklin (Gladstone Institutes, San Francisco, USA) for providing the hiPSCs and technical advice on the cell line, and Dr. Chenang Lyu for technical assistance with the isochoric chamber assembly process.
© 2021, The Author(s).
PubMed: MeSH publication types
- Journal Article
- Research Support, U.S. Gov't, Non-P.H.S.
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- 1 Active
ATP-Bio: NSF Engineering Research Center for Advanced Technologies for the Preservation of Biological Systems (ATP-Bio)
9/1/20 → 8/31/25
Project: Research project