Abstract
3D printing was used to develop an open access device capable of simultaneous electrical and mechanical stimulation of human induced pluripotent stem cells in 6-well plates. The device was designed using Computer-Aided Design (CAD) and 3D printed with autoclavable, FDA-approved materials. The compact design of the device and materials selection allows for its use inside cell incubators working at high humidity without the risk of overheating or corrosion. Mechanical stimulation of cells was carried out through the cyclic deflection of flexible, translucent silicone membranes by means of a vacuum-controlled, open-access device. A rhythmic stimulation cycle was programmed to create a more physiologically relevant in vitro model. This mechanical stimulation was coupled and synchronized with in situ electrical stimuli. We assessed the capabilities of our device to support cardiac myocytes derived from human induced pluripotent stem cells, confirming that cells cultured under electromechanical stimulation presented a defined/mature cardiomyocyte phenotype. This 3D printed device provides a unique high-throughput in vitro system that combines both mechanical and electrical stimulation, and as such, we foresee it finding applications in the study of any electrically responsive tissue such as muscles and nerves.
Highlights
Recent advances in tissue engineering, along with the discovery of induced pluripotent stem cells has opened the doors to new therapies for treating cardiovascular diseases (CVD)[1,2]
The device was designed using Computer-Aided Design (CAD) software (SolidWorks, Dassault Systems) and 3D printed in an Ultimaker S5 with Nylon 680 filament (Fig. 1A)
The current body of literature speaks to the clinical value of human-induced pluripotent stem cells as therapeutic agents for tissue regeneration and in vitro drug screening
Summary
Recent advances in tissue engineering, along with the discovery of induced pluripotent stem cells (iPSC) has opened the doors to new therapies for treating cardiovascular diseases (CVD)[1,2]. Researchers have developed and improved differentiation techniques in order to obtain cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CM)[3,4] These cells have been used both as therapeutics for the regeneration of lost cardiac tissue as well as in in vitro models of cardiac tissues for drug screening[1,5,6,7]. They found that after two weeks of static stress followed by one week of electrical pacing, cell alignment, cardiac hypertrophy, passive stiffness, and contractility was improved Effectively incorporating both stimulation regimes, these devices, as well as those developed by other research groups[2,5,28,29], have shown great variability in the results of the maturation of hiPSC-CMs, determined by protein and gene expression, cell alignment, and electrical conductivity. We set out to develop a readily accessible and cost-effective device able to electro-mechanically stimulate cells, which we foresee being instrumental in the harmonization of cell differentiation protocols amongst laboratories as well as the development of models for the study of stem cell therapies and use in drug screening
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