As the field of cardiac and skeletal muscle tissue engineering expands, so does the need for accurate and reliable systems to generate in vitro 3D tissues and analyze their functional properties.... Show moreAs the field of cardiac and skeletal muscle tissue engineering expands, so does the need for accurate and reliable systems to generate in vitro 3D tissues and analyze their functional properties. In this study, the Cuore is introduced, a system that integrates sensors based on optical fibers and uses the principle of light interferometry to detect the contraction of 3D Tissue Engineered Skeletal Muscles (3D-TESMs). The technology employed in the Cuore allows for reproducible and multichannel force measurements down to a nano-Newtons resolution while maintaining sterility and permitting continuous non-invasive recording within and outside standard tissue culture incubators. Thanks to the integrated electrodes for electrical pulse stimulation (EPS), 3D-TESMs generated from three independent hiPSC-derived myogenic progenitors (MPs) lines are stimulated and the contractility is recorded over the course of a week. Through the modulation of different EPS parameters, the optimal combination to induce the 3D-TESMs in producing fully fused tetani without causing damage is determined. Furthermore, 3D-TESMs from different lines exhibit characteristic signatures of spontaneous contractility and response to caffeine, verapamil, and the & beta;-agonist clenbuterol. The ease of use, high sensitivity, and the integrated electrodes and sensors make the Cuore an ideal technology to investigate the biology of contractile tissues and their response to drugs.The expanding field of cardiac and skeletal muscle tissue engineering demands efficient solutions to study the biology of the tissues and model disorders. In this study the Cuore system is presented, employing optical fiber sensors and light interferometry to detect 3D Tissue Engineered Skeletal Muscle contractions. Thanks to nano-Newton resolution force measurements and integrated electrical pulse stimulation, contractility of hiPSC-derived tissues is evaluated. Additionally, its efficacy in assessing drug responses, like caffeine and clenbuterol, showcases its potential in studying contractile tissue biology.image Show less
Iuliano, A.; Wal, E. van der; Ruijmbeek, C.W.B.; in't Groen, S.L.M.; Pijnappel, W.W.M.P.; Greef, J.C. de; Saggiomo, V. 2020
The transition from 2D to 3D engineered tissue cultures is changing the way biologists can perform in vitro functional studies. However, there has been a paucity in the establishment of methods... Show moreThe transition from 2D to 3D engineered tissue cultures is changing the way biologists can perform in vitro functional studies. However, there has been a paucity in the establishment of methods required for the generation of microdevices and cost-effective scaling up. To date, approaches including multistep photolithography, milling and 3D printing have been used that involve specialized and expensive equipment or time-consuming steps with variable success. Here, a fabrication pipeline is presented based on affordable off-the-shelf 3D printers and novel replica molding strategies for rapid and easy in-house production of hundreds of 3D culture devices per day, with customizable size and geometry. This pipeline is applied to generate tissue engineered skeletal muscles in vitro using human induced pluripotent stem cell-derived myogenic progenitors. These production methods can be employed in any standard biomedical laboratory. Show less
