Deciphering Role of Wnt Signalling in Cardiac Mesoderm and Cardiomyocyte Differentiation from Human iPSCs: Four-dimensional control of Wnt pathway for hiPSC-CMs differentiation

摘要: Differentiation of cardiomyocytes (CMs) from human induced pluripotent stem cells (hiPSCs) is critically dependent upon the regulation Wnt signaling pathway. The mechanisms remain unclear with regard to dose and timing each differentiation inducer, interaction inducers that regulate in mesendoderm specification cardiac mesoderm. Consequently, it remains far optimal efficiency consistency hiPSC lines CMs. Here, we have carefully deciphered role pathway manipulation on mesoderm a dosage time manner. To examine hypothesis fate hiPSC-CMs dictated by temporal spatial factors Wnt, evaluate hiPSC-CM with: (1) two-phase modulation (2) variant GSK3β inhibitors, (3) treatment insulin, (4) 3-dimentional suspension culture environment iPSC-CM differentiation. results highlight importance mesoderm, which needs precisely on/off This dictates final purity derived cardiomyocytes. After initial activation generate mesendoderm, maintenance at an appropriate critical direct cell into Otherwise, lower signals lead definitive endoderm higher induce presomitic not only greater than 90% cTnT+ but also high yield under both monolayer conditions. Thus, current findings provide insights decipher mechanism differentiation, more importantly guidelines for consistent high-yield high-quality production cardiovascular research.