(W1165) Efficient Production of Cardiomyocytes in 2D en 3D: Wnt and Insulin Enhance Proliferation and Inhibit Maturation of Human iPSC-derived Cardiomyocytes via TCF and FOXO Signaling
Cardiologist / Assistant Professor Vrije Universiteit Amsterdam, Noord-Holland, Netherlands
Abstract: Embryonic signaling pathways exert stage-specific effects during cardiac development, yet the precise signals for proliferation or maturation remain elusive. To uncover the cues for proliferation, we performed a combinatory cell cycle screen for Insulin and glycogen synthase kinase-3 (GSK-3) inhibition in spontaneously beating human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).
Our analysis for proliferation, and subsequential downstream sarcomere development, gene expression analysis and molecular interventions identified a temporal interplay between Insulin/Akt/FOXO and CHIR99021/Wnt/GSK-3/TCF signaling. Combined pathway activation led to proliferation of immature hiPSC-CMs with low sarcomere and mitochondria content, while in the absence of pathway activators, cardiomyocytes rapidly exited the cell cycle and fetched higher organization of sarcomeres and mitochondria.
Our data demonstrates two important pathways which enhance proliferation and inhibit maturation, and provides molecular mechanistic understanding of these cell fate decisions in immature hiPSC-CMs. Moreover, this technology allows for expansion of hiPSC-CMs 2-dimensional (2D) large culture flasks and 3-dimensional (3D) stirrer flasks for the production of hundreds of millions of cells. This facilitates a desired scale up for drug testing and large heart tissue generation.
