Sama Salih
Research Mentor(s): Dr. Todd Herron
Department or Program: Frankel Cardiovascular Regeneration Core Laboratory
Authors: Todd J. Herron, Prakaimuk Saraithong, Devika Baddhan, Hailey Cunningham
Session: Session 1: 12:00pm-12:50pm
Poster: 41
Abstract
Conventional cell culture processing procedures do not support successful cryopreservation and functional recovery of human stem cell-derived cardiomyocytes. Cryopreservation is crucial for human cardiomyocyte production and downstream workflows as it facilitates long-term storage and distribution to other labs and institutions. . Here we utilized an innovative Artificial Intelligence guided laser cell processing approach to produce stem cell-derived cardiomyocytes. We tested the hypothesis that AI-laser-processed cardiomyocytes will recover from liquid nitrogen cryopreservation and yield functionally contracting cells with normal drug responsiveness. Cardiomyocytes were purified using an AI-guided laser system (Kataoka), which precisely targeted and eliminated non-cardiomyocytes. The enriched cardiomyocytes were subsequently collected and cryopreserved using commercially available solutions, then stored in liquid nitrogen for 1 week prior to thaw and functional analysis. Our findings showed that frozen cardiomyocytes retained high viability with minimal cell death. The spontaneous beat rate of thawed cardiomyocytes was measured using a microplate reader and imager (Agilent BioTek Gen5). Both atrial and ventricular cardiomyocytes were evaluated, with both CMs subtypes exhibiting the expected spontaneous beat rates. The cardiomyocytes responded appropriately to two pharmacological agents with well-known cardiac effects: Azimilide for atrial cells and isoprenaline for ventricular cells. Azimilide, an anti-arrhythmic agent with potassium channel blocking effects, modulated the electrophysiological properties of atrial CMs and slowed the beat rate, while isoprenaline, a β-adrenergic agonist, increased the contraction rate in ventricular cells. This study underscores the benefits of AI-guided laser purification for cryopreserved cardiomyocytes, demonstrating that this method not only preserves cell viability and functionality but also enhances overall cell quality. The ability of cryopreserved CMs to retain their structural, metabolic, and electrophysiological properties post-thaw makes them a valuable resource for high-throughput diagnostics, personalized medication screening, and cardiac regeneration research. This approach could transform the cell culturing pipeline, increasing the availability and reliability of viable cardiomyocytes for various experimental and therapeutic applications.
