Protocol for the Generation of Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

This protocol offers a detailed procedure for the in vitro differentiation of human pluripotent stem cells (hPSCs) to multipotent hematopoietic progenitors that arise from SOX17+ hemogenic endothelium, mimicking intra-embryonic, HOXA-positive, aorta-gonad mesonephros (AGM) hematopoiesis. The generat...

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Bibliographic Details
Published in:	STAR protocols Vol. 1; no. 3; p. 100130
Main Authors:	Nafria, Monica, Bonifer, Constanze, Stanley, Edouard Guy, Ng, Elizabeth Siewsun, Elefanty, Andrew George
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 18-12-2020 Elsevier
Subjects:	Cell Culture Techniques - methods Cell Differentiation - drug effects Cell Differentiation - physiology Gonads - cytology Hematopoiesis - physiology Hematopoietic Stem Cells - cytology Hematopoietic Stem Cells - metabolism Humans Mesonephros - cytology Models, Biological Pluripotent Stem Cells - metabolism Pluripotent Stem Cells - physiology Protocol
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Summary:	This protocol offers a detailed procedure for the in vitro differentiation of human pluripotent stem cells (hPSCs) to multipotent hematopoietic progenitors that arise from SOX17+ hemogenic endothelium, mimicking intra-embryonic, HOXA-positive, aorta-gonad mesonephros (AGM) hematopoiesis. The generated endothelium displays transcriptional similarities to cells sorted from human 5-week AGM, and CD45+CD34+RUNX1C+ progenitors share an accessible chromatin profile with adult hematopoietic stem cells and multipotent progenitors. Therefore, this protocol is suitable for the mechanistic study of human multipotent progenitor development and for modeling childhood leukemias. For complete details on the use and execution of this protocol, please refer to Nafria et al. (2020). [Display omitted] For a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.xpro.2020.100130. •Protocol generating definitive hematopoiesis from human pluripotent stem cells•Mimics intra-embryonic hematopoiesis from HOXA+SOX17+ hemogenic endothelium•Generates blood progenitors resembling adult HSCs and multipotent progenitors•Enables the study of human hematopoiesis under unperturbed and leukemic conditions This protocol offers a detailed procedure for the in vitro differentiation of human pluripotent stem cells (hPSCs) to multipotent hematopoietic progenitors that arise from SOX17+ hemogenic endothelium, mimicking intra-embryonic, HOXA-positive, aorta-gonad mesonephros (AGM) hematopoiesis. The generated endothelium displays transcriptional similarities to cells sorted from human 5-week AGM, and CD45+CD34+RUNX1C+ progenitors share an accessible chromatin profile with adult hematopoietic stem cells and multipotent progenitors. Therefore, this protocol is suitable for the mechanistic study of human multipotent progenitor development and for modeling childhood leukemias.
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ISSN:	2666-1667 2666-1667
DOI:	10.1016/j.xpro.2020.100130