show Abstracthide AbstractRegulation of gene expression acts at numerous complementary levels to control and refine the protein abundance. The analysis of mRNAs associated to polysomes, called polysome profiling, has been used to investigate post-transcriptional mechanisms involved in different biological processes. Pluripotent stem cells, as human embryonic stem cells (hESCs), are able to differentiate into a variety of cell lineages and the cell commitment progression is carefully orchestrated. Genome-wide expression profiling had provided the possibility to investigate transcriptional changes during cardiomyogenic differentiation, however, a more accurate study regarding post-transcription regulation is required. In the present work, we isolated and high-throughput sequenced ribosome-free and polysome-bound RNAs from undifferentiated pluripotent stem cells and the subsequent differentiation stages of cardiomyogenesis: embryoid body aggregation, mesoderm, cardiac progenitor and cardiomyocytes. Expression of developmental markers were followed by flow cytometry and quality analysis were performed as technical controls to ensure high quality data. Our dataset provides valuable information about hESC cardiac differentiation and can be used to investigate genes potentially controlled by post-transcriptional mechanisms.