Cellular functions are frequently exploited as processing components for integrated chemical systems such as biochemical reactors and bioassay systems. Here, we have created a new cell-based microsystem exploiting the intrinsic pulsatile mechanical functions of cardiomyocytes to build a cellular micropump on-chip using cardiomyocyte sheets as prototype bio-microactuators. We first demonstrate cell-based control of fluid motion in a model microchannel without check valves and evaluate the potential performance of the bio-actuation. For this purpose, a poly(dimethylsiloxane) (PDMS) microchip with a microchannel equipped with a diaphragm and a push-bar structure capable of harnessing collective cell fluid mechanical forces was coupled to a cultured pulsating cardiomyocyte sheet, activating cell-based fluid movement in the microchannel by actuating the diaphragm. Cell oscillation frequency and correlated fluid displacement in this system depended on temperature. When culture temperature was increased, collective cell contraction frequency remained cooperative and synchronous but increased, while displacement was slightly reduced. We then demonstrated directional fluid pumping within microchannels using cantilever-type micro-check valves made of polyimide. A directional flow rate of nL min(-1) was produced. This cell micropump system could be further developed as a self-actuated and efficient mechanochemical transducer requiring no external energy sources for various purposes in the future.

Natural cellular functions are increasingly exploited for integrated chemical systems such as biochemical reactors and biosensors. We propose to utilize the intrinsic mechanical function of cardiomyocytes, converting chemical energy into mechanical energy. In this report, we demonstrate the working principle of our proposed poly(dimethylsiloxane) (PDMS) based cardiomyocyte bio-microactuator using fabricated PDMS micropillars driven to repetitive motion by attached pulsating cardiomyocytes. Sheets of PDMS embedded with an array of micropillars were fabricated and modified for cardiomyocyte attachment in culture. Primary neonatal rat cardiomyocytes were cultured on the array, attaching to the micropillars and substratum successfully, and exhibiting their typical spontaneous, pulsatile phenotype. Micropillars beat with the coupled cells spontaneously without any triggers. The beat frequency was 1.4 Hz at 37 degrees C and the displacement of the top of the pillar that beat most strongly in our observation was 2.8+/-0.2 microm. From this result, contractile forces of cultured cardiomyocytes were estimated to exceed 3.5 microN. The estimated force is far greater than that of a previously described hydrogel-based cardiomyocyte bio-microactuator (K. Morishima et al., in Micro Total Analysis Systems 2003, ed. M. A. Northrup et al., The Transducers Research Foundation, San Diego, CA, vol. 2, pp. 1125-1128). PDMS compatibility as a base material for bio-microactuator design using cultured cardiomyocytes was verified. This PDMS-based cell microactuator worked for about one week without exchange of the culture medium, and this system could be developed for various purposes in the future as self-actuated and efficient mechanochemical transducers without external energy source requirements.

Despite attempts in a number of studies to utilize muscle tissue and cells as microactuators, all of the biohybrid microdevices have been operable only in the culture medium and none have worked in air due to the dry environment. This paper demonstrates an atmospheric-operable bioactuator (AOB) fabricated by packaging an insect dorsal vessel (DV) tissue with a small amount of culture medium inside a capsule. The AOB, consisting of microtweezers and the capsule, was designed based on a structural simulation that took into account the capillary effect. The base part of the microtweezers was deformed by spontaneous contractions of the DV tissue in the medium inside the capsule, by which the front edges of the microtweezer arms projecting above the medium surface were also deformed. First, we confirmed in the medium that the DV tissue was able to reduce the gap between the arm tips of the microtweezers. After taking the AOB out of the medium, as we expected, the AOB continued to work in air at room temperature. The gap reduction in air became larger than the one in the medium due to a surface tension effect, which was consistent with the simulation findings on the surface tension by the phase-field method. Second, we demonstrated that the AOB deformed a thin-wall ring placed between its tips in air. Third, we measured the lifetime of the AOB. The AOB kept working for around 40 minutes in air, but eventually stopped due to medium evaporation. As the evaporation progressed, the microtweezers were pressed onto the capsule wall by the surface tension and opening and closing stopped. Finally, we attempted to prevent the medium from evaporating by pouring liquid paraffin (l-paraffin) over the medium after lipophilic coating of the capsule. As a result, we succeeded in prolonging the AOB lifetime to more than five days. In this study, we demonstrated the significant potential of insect muscle tissue and cells as a bioactuator in air and at room temperature. By integrating insect tissue and cells not only into a microspace but also onto a substrate, we expect to realize a biohybrid MEMS device with various functions in the near future.