Single cell transcriptomics of hiPSC-derived forward programmed megakaryocytes

We have published a novel method for the large-scale in vitro generation of megakaryocytes (MKs), the blood platelet precursors, by applying a transcription factor (TF) driven forward programming strategy to human pluripotent stem cells (hPSCs). We have demonstrated that the concurrent expression of GATA1, FLI1 and TAL1 in hPSCs and chemically defined culture conditions with minimal supportive cytokines produce highly pure MK cultures with long-term growth and release of functional platelets. Unravelling the molecular mechanisms underlying MK forward programming will bring biological insights that shall lead to improvements of the MK programming technology. Particularly, we have been focusing on the characterisation of the MK progenitor that allows long-term expansion of pure MK cultures, which is a key asset of the method. Using single cell RNA sequencing of long-term cultures, we have started to identify and functionally confirmed surface markers of the MK progenitor. Whole genome single cell transcriptome analysis of long-term MK culture (one hiPSC line), balanced for the eight transgenic patterns (upstream FACS using transgene expression reporters)