Cardiovascular disease is the world’s killer number one and despite new therapies, the number of heart failure is expected to increase further in the coming decades. One of the key reasons underlying this high number is the extremely low regeneration capacity of mammalian cardiomyocytes. To boost this very low, but existing endogenous cardiomyocyte proliferation potential is a major focus in regenerative medicine. However, in the absence of a suitable human model system, screening of potential drug candidates promoting regeneration is limited. Therefore, we have established a human induced pluripotent stem cell (hiPSC)-derived cardiomyocyte model for live-cell monitoring of cardiomyocytes proliferation by employing a fluorescent reporter gene. In detail, we generated a stable hiPSC cell line using a lentiviral vector harboring the constitutive promoter of the human phosphoglycerate kinase (hPGK) driving a cell-cycle reporter gene. This consists of the N-terminal region of Cyclin B1 including a cell-cycle regulated destruction signal, fused to eGFP (CycB1-eGFP), which can efficiently label replicating cells in the S/G2/M phases of the cell cycle. We first demonstrated that the transgene expression is stable for over ten passages, that the hiPSCs remained pluripotent and can be readily differentiated into cardiomyocytes. We observed that around 3% of the differentiated cardiomyocytes were eGFP+. This was in contrast to around 12% Ki67+ (proliferation marker) cells, speculating that the CycB1-eGFP reporter gene might label faithfully proliferating CMs whereas Ki67 is only a marker for the cell cycle activity. We sorted cardiomyocytes into eGFP+ (proliferating) and eGFP- (non-proliferating) fractions and performed whole transcriptome RNA sequencing and gene set enrichment analysis (GSEA). As expected eGFP+ cardiomyocytes show a significant upregulation of genes involved in DNA replication and cell cycle progression. Interestingly, we found that the expression of metabolic programs and structural genes of proliferating cardiomyocytes exhibit a molecular signature similar to embryonic/neonatal cardiomyocytes, which are known to possess proliferative properties. Treatment of our cardiomyocyte reporter cells with the Wnt pathway activator CHIR99021 led to a significant increase in eGFP+ cardiomyocytes, demonstrating that our reporter gene faithfully marks cardiomyocytes through cell cycle progression and mitosis. Time-lapse live imaging of eGFP+ cardiomyocytes further demonstrated that our reporter construct labels cell division and not endomitosis of cardiomyocytes, confirming the reliability of our reporter system for live-cell tracking of proliferating cardiomyocytes. We proposed that our human cardiomyocytes reporter cell line represents an innovative translational platform for large-scale screening of pro-regenerative compounds that in the future could be used to trigger cardiac repair by stimulating endogenous proliferation of pre-existing cardiomyocytes.