Extended Data Figure 6: Identification of the source of engraftable cells within the HE-7TF population.

a, Conversion of haemogenic endothelium (HE) into haematopoietic stem/progenitor cells by seven transcription factors requires EHT. Haemogenic endothelium or haemogenic endothelium grown in EHT medium for 3 days was transduced with seven transcription factors and transplanted into mice followed by bone marrow analysis at 4 weeks. Cells grown after 3 days of EHT showed CD45⁺ cells while those that were not grown under EHT conditions did not show CD45⁺ cells. b, Human umbilical vein endothelial cells were transduced with seven transcription factors or GFP lentiviral vectors, then cultured in EHT medium with Dox for a week. Flow cytometry analysis of PECAM (EC marker) and CD34 (haematopoietic marker) is shown. Human umbilical vein endothelial cells transduced with seven transcription factors fails to produce robust CD34⁺ cell population. c, Twenty-five thousand CD34⁺CD43⁺CD45⁺ (triple positive, TP) or CD34⁺CD43⁻CD45⁻ (single positive, SP) cells were FACS-isolated and transplanted (N = 5 mice per group). Engraftment of human CD45⁺ cells was assessed in peripheral blood (PB) at 8 weeks. FACS plots of human CD45 and mouse CD45.1 of TP (left) and single positive (right) transplanted mice at the 8 week time point are shown. d, Multi-lineage engraftment of bone marrow and spleen from primary recipient mouse at 6 weeks is shown. CD34⁺CD43⁺CD45⁺ cells were intravenously injected.

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