Transgene-free haematopoietic stem and progenitor cells from human induced pluripotent stem cells

Despite considerable efforts, the generation of vector-free hematopoietic stem cells (HSCs) from human induced pluripotent stem cells (hiPSCs) is not yet possible, preventing the development of cell therapies to cure hematologic diseases. Here, we report the generation of HSCs with multilineage and self-renewal potential from hiPSCs using a specific one-step procedure over a culture period of 17 days. After injection into immunocompromised mice, cells derived from hiPSCs settle in the bone marrow and form a robust multicellular hematopoietic population that can be serially transplanted. RNA sequencing of individual cells suggests that this repopulating activity likely relies on a hematopoietic population transcriptionally similar to HSCs derived from human embryonic aorta. Overall, our results should enable the generation of HSCs from hiPSC and will be instrumental to identify important regulators of HSC production during human ontogeny. Embroid bodies (EB) derived from hiPS and cultivated until the chosen timepoints were dissociated in single cells and analysed by scRNAseq using the Chromium 10X v2 technology. Mouse bone marrow 20 weeks after engraftment of the EB cells was flushed and analysed by scRNAseq using the Chromium 10X v3.1 technology