Generation of a human induced pluripotent stem cell reporter line to investigate cell division and proliferation.

Understanding cell division in disease contexts is of paramount importance for elucidating disease mechanisms and developing regenerative therapies, such as cardiac regeneration. Nevertheless, tools for identifying and to studying key factors in regenerative processes in human cells remain scarce. Here, we generated a human induced pluripotent stem cell (hiPSC) reporter line expressing a cell cycle-regulated cyclinB1-eGFP construct that enables live tracking of proliferating human cells. The reporter hiPSC line successfully differentiated into cardiomyocytes (CMs), endothelial cells (ECs), and fibroblasts (FBs), with eGFP+ cells identifying actively dividing populations across lineages. Each cell type exhibited appropriate lineage-specific marker expression and high differentiation efficiency. Importantly, the cyclinB1-eGFP system allowed real-time identification and tracking of proliferating (eGFP+) cells within these differentiated populations. This tool provides an innovative platform for screening potential pro-proliferative compounds, facilitating the discovery of novel therapies to stimulate or inhibit cell division.