Abstract
We have developed our original tissue engineering technology “cell sheet engineering” utilizing temperature-responsive culture dishes. The cells are confluently grown on a temperature-responsive culture dish and can be harvested as a cell sheet by lowering temperature without enzymatic digestion. Cell sheets are high-cell-density tissues similar to actual living tissues, maintaining their structure and function. Based on this “cell sheet engineering”, we are trying to create functional cardiac tissues from human induced pluripotent stem cells, for regenerative therapy and in vitro drug testing. Toward this purpose, it is necessary to evaluate the contractility of engineered cardiac cell sheets. Therefore, in the present study, we developed a contractile force measurement system and evaluated the contractility of human iPSC-derived cardiac cell sheet-tissues. By attaching the cardiac cell sheets on fibrin gel sheets, we created dynamically beating cardiac cell sheet-tissues. They were mounted to the force measurement system and the contractile force was measured stably and clearly. The absolute values of contractile force were around 1 mN, and the mean force value per cross-sectional area was 3.3 mN/mm2. These values are equivalent to or larger than many previously reported values, indicating the functionality of our engineered cardiac cell sheets. We also confirmed that both the contractile force and beating rate were significantly increased by the administration of adrenaline, which are the physiologically relevant responses for cardiac tissues. In conclusion, the force measurement system developed in the present study is valuable for the evaluation of engineered cardiac cell sheet-tissues, and for in vitro drug testing as well.
Highlights
Recent advances in tissue engineering are greatly promoting its application to regenerative therapies, in vitro drug testing, and pathological investigations
After the cell sheet transfer procedure, we confirmed that the cardiac cell sheet was attached on the surface of a fibrin gel sheet by phasecontrast microscopy (Fig 4C) and hematoxylin and eosin (H&E) staining of paraffin sections (Fig 4D)
In order to evaluate the contractile properties of engineered cardiac cell sheet-tissues with measurable quantities, in the present study we developed a system to measure the contractile forces of cardiac cell sheet-tissues
Summary
Recent advances in tissue engineering are greatly promoting its application to regenerative therapies, in vitro drug testing, and pathological investigations. One of the most widespread methodologies in tissue engineering is to mix cells with a biocompatible scaffold of natural and/or synthetic polymers such as collagen gel, poly(lactide-co-glycolide), and so on [1, 2]. Contractile force measurement of human iPSC-derived cardiac cell sheet-tissue.
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