Electrical Stimulation For The Maturation Of Induced Pluripotent Stem Cell-derived Cardiomyocytes

Heart disease is the leading cause of death in the U.S. and worldwide. Cardiac tissue engineering offers promising diagnostic tools and treatments. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are an attractive cell source for these tissue engineering applications due to their patient-specificity and theoretical capacity to supply limitless cell quantities. However, these hiPSC-CMs are immature compared to healthy, adult cardiomyocytes, demonstrating immature morphology, metabolics, and electrophysiology. Previous research has shown electrical stimulation shows promise in maturation of hiPSC-CMs. We aim to create an in vitro platform for the electrophysiological maturation of hiPSC-CMs using applied electrical stimulation during the differentiation process. We designed and fabricated a bioreactor for the application of electrical stimulation to hiPSC-CMs in culture. The bioreactor was created by laser cutting two parallel slits into the lid of a 12-well tissue culture plate, through which conductive Toray paper was placed. The space remaining between the slits and the Toray paper was sealed with polydimethylsiloxane (PDMS). Alligator clips were used to connect a function generator to the Toray paper. When sterilized with 70% ethanol, this modified 12-well plate lid can be used to apply electrical stimulation to cells cultured in a 12-well plate, as shown in Figure 1. Future work includes the application of electrical stimulation on day 3 of a well-established Wnti-directed monolayer protocol for the differentiation of hiPSCs into hiPSC-CMs. We expect to observe a resultant improvement in iPSC-CM’s genetic and electrophysiological maturation.