Practical adoption of state-of-the-art hiPSC-cardiomyocyte differentiation techniques.

摘要: Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are a valuable resource for cardiac therapeutic development; however, generation of these cells in large numbers and high purity is limitation widespread adoption. Here, design experiments (DOE) used to investigate the differentiation space three hiPSC lines when varying CHIR99027 concentration seeding density, novel image analysis developed evaluate plate coverage initiating differentiation. Metabolic selection via lactate purifies hiPSC-cardiomyocyte populations, bioenergetic phenotype engineered tissue mechanics purified unpurified hiPSC-cardiomyocytes compared. Findings demonstrate that one day after plating, low (3 μM) Chiron 72 x 103 cells/cm2 density result peak (50-90%) all lines. Our results confirm metabolic with shifts metabolism towards oxidative phosphorylation, but this more "mature" does not by itself mature contractile tissues at week culture 3D tissues. This study provides widely adaptable methods including code parameters refining describes practical implications downstream engineering applications.