Measuring the Contractile Force of Multilayered Human Cardiac Cell Sheets
Publication: Tissue Engineering Part C: Methods
Volume 26, Issue Number 9
Abstract
Three-dimensional (3D) cardiac tissue reconstruction using tissue engineering technology is a rapidly growing area of regenerative medicine and drug screening development. However, there remains an urgent need for the development of a method capable of accurately measuring the contractile force of physiologically relevant 3D myocardial tissues to facilitate the prediction of human heart tissue drug sensitivity. To this end, our laboratory has developed a novel drug screening model that measures the contractile force of cardiac cell sheets prepared using temperature-responsive culture dishes. To circumvent the difficulties that commonly arise during the stacking of cardiomyocyte sheets, we established a stacking method using centrifugal force, making it possible to measure 3D myocardial tissue. Human induced pluripotent stem cell-derived cardiomyocytes were seeded in a temperature-responsive culture dish and processed into a sheet. The cardiac cell sheets were multilayered to construct 3D cardiac tissue. Measurement of the contractile force and cross-sectional area of the multilayered 3D cardiac tissue were then obtained and used to determine the relationship between the cross-sectional area of the cardiac tissue and its contractile force. The contractile force of the 1-, 3-, and 5-layer tissues increased linearly in proportion to the cross-sectional area. A result of 6.4 mN/mm2, accounting for one-seventh of the contractile force found in adult tissue, was obtained. However, with 7-layer tissues, there was a sudden drop in the contractile force, possibly because of limited oxygen and nutrient supply. In conclusion, we established a method wherein the thickness of the cell sheets was controlled through layering, thus enabling accurate evaluation of the cardiac contractile function. This method may enable comparisons with living heart tissue while providing information applicable to regenerative medicine and drug screening models.
Abstract
Impact statement
Cell sheet technology was used to construct myocardial tissues to visually confirm pulsation. By stacking these cell sheets, a three-dimensional (3D) myocardial tissue was created and a method for measuring the contractile force was constructed. Our results revealed that the contraction force of 1- to 5-layer sheets increased linearly in tandem with the cross-sectional area, giving rise to the synchronized beats in the 3D myocardial tissue. This method allows for a more accurate evaluation of myocardial contractile force.
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Information & Authors
Information
Published In
Tissue Engineering Part C: Methods
Volume 26 • Issue Number 9 • September 2020
Pages: 485 - 492
PubMed: 32799760
Copyright
Copyright 2020, Mary Ann Liebert, Inc., publishers.
History
Published online: 17 September 2020
Published in print: September 2020
Published ahead of production: 16 August 2020
Accepted: 11 August 2020
Received: 28 June 2020
Topics
Authors
Authors' Contributions
K.S. designed and performed the experiments, analyzed the data, and wrote the article. H.T. designed and performed the experiments and analyzed the data. Y.T. performed the experiments and analyzed the data. D.S. designed and performed the experiments. K.M. designed and performed the experiments and supervised the project. K.I. designed the experiments and supervised the project. T.S. designed the experiments, analyzed the data, wrote the article, and supervised the project. M.U. designed the experiments and supervised the project.
Disclosure Statement
T.S. was a member of the scientific advisory board and a shareholder of CellSeed, Inc. (Tokyo, Japan). Tokyo Women's Medical University receives research funds from CellSeed, Inc.
T.S. and K.M. are inventors of the bioreactor systems for iPSC proliferation and cardiomyocyte differentiation. Tokyo Women's Medical University, T.S., and K.M. received a license fee from ABLE Corporation, Tokyo, Japan.
T.S., D.S., and K.M. are inventors of the contractile force measurement system of Nihon Kohden Corporation, Tokyo, Japan. Tokyo Women's Medical University receives research funds from Nihon Kohden Corporation.
Funding Information
This research was supported by Japan Agency for Medical Research and Development under grant number JP18he0702249.
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