Proteomic Approach Using DIA-MS Identifies Morphogenesis-Associated\nProteins during Cardiac Differentiation of Human iPS Cells

Human-induced pluripotent\nstem cell (hiPSC)-derived cardiomyocytes\nhave potential applications in regenerative medicine. The quality\nby design (QbD) approach enables the efficiency and quality assurance\nin the manufacturing of hiPSC-derived products. It requires a molecular\nunderstanding of hiPSC differentiation throughout the differentiation\nprocess; however, information on cardiac differentiation remains limited.\nProteins associated with the early stages of cardiac differentiation\nwould be useful in the cardiomyocyte quality assessment. Here, we\nperformed quantitative proteomics of hiPSC intermediate cells in the\nearly phase of cardiac differentiation to better understand their\nmolecular characteristics. Proteomic profiles suggested that day 5–7\ncells were in the morphogenetic stage of cardiac differentiation.\nTrophoblast glycoprotein (TPBG) was the most up-regulated protein\nin the morphogenetic stage; it was previously shown to be up-regulated\nduring differentiation into neural stem cells. Proteomics of TPBG-knockdown\ncells revealed that TPBG is involved in cell proliferation and is\nrelated to the cardiomyocyte yield, suggesting that it could be used\nas a marker in QbD development. Our approach helps us understand the\nmolecular basis of hiPSC differentiation and could be a powerful tool\nin QbD-based manufacturing.