The differentiation of megakaryocytes from human pluripotent stem cells in vitro offers intriguing new perspectives for research and transfusion medicine. However, applications have been hampered by the low efficiency of cytokine driven differentiation protocols leading to poor megakaryocyte purity and yield. Here we describe a novel forward programming approach relying on the combined ectopic expression of the three transcription factors GATA1, FLI1, and TAL1 in human pluripotent stem cells for large scale production of mature megakaryocytes using chemically defined culture and minimum cytokines.