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Here we propose a room temperature operable muscle powered bioactuator by dorsal vessel (tubular insect heart) cells. Insect cells are generally robust over a range of culture conditions compared to mammalian cells. In this study, we conducted two experiments to show the potential of the insect cell as an actuator. First, beating frequency and beating force of dorsal vessels as both tissues and cells were measured to evaluate their performance. In the experiment, the ability to control the dorsal vessel cells confirmed by the chemical dosage. It was succeeded in the measurement of the driving force of the dorsal vessel. Second, two types of cell sheet were fabricated using poly(dimethylsiloxane) PDMS thin film and agarose gel. Both types of cell sheet were successfully detached from cell culture surface and contracted spontaneously. These results showed the possibility of a bioactuator powered by a dorsal vessel cell sheet.