Human pluripotent stem cell derived hematopoietic cells using expanded potential stem cell media

We recently developed a novel chemical media formulation, Expanded Potential Stem Cell Medium (EPSCM), that maintains mouse pluripotent stem cells (PSCs) in a unique cellular state that affords extraembryonic differentiation capacity. Here, we have used EPSCM to stably maintain human PSCs and used EPSCM-maintained iPSCs (EC-EPSCM) to study in vitro hematopoietic differentiation by embryoid body (EB) formation. We found that human PSCs maintained in EPSCM can spontaneously form embryoid bodies and efficiently undergo in vitro hematopoiesis using a simple differentiation protocol. EC-EPSCM PSCs generated primitive-wave and definitive-wave hematopoietic cells, which were chracterized by RNA-sequencing analysis. Overall design: Characterization of four hematopoietic cell populations (two cell types from two different time points) generated by the embyoid body (EB) differentiation of EPSCM-maintained human iPSC line (EC-EPSCM), which represent primitive-wave hematopoietic cells and definitive-wave hematopoietic cells. Cell populations were purified by fluorescence-activated cell sorting (50-cells per sample) and analyzed by RNA-sequencing using the Smartseq2 protocol.