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

Monica Nafria; Constanze Bonifer; Edouard Guy Stanley; Elizabeth Siewsun Ng; Andrew George Elefanty

doi:10.1016/j.xpro.2020.100130

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

Monica Nafria
, Constanze Bonifer
, Edouard Guy Stanley
, Elizabeth Siewsun Ng^*
, Andrew George Elefanty^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

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).

Original language	English
Article number	100130
Number of pages	25
Journal	STAR Protocols
Volume	1
Issue number	3
Early online date	16 Oct 2020
DOIs	https://doi.org/10.1016/j.xpro.2020.100130
Publication status	Published - 18 Dec 2020

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title = "Protocol for the Generation of Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells",

abstract = "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).",

author = "Monica Nafria and Constanze Bonifer and Stanley, \{Edouard Guy\} and Ng, \{Elizabeth Siewsun\} and Elefanty, \{Andrew George\}",

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AU - Nafria, Monica

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AU - Stanley, Edouard Guy

AU - Ng, Elizabeth Siewsun

AU - Elefanty, Andrew George

PY - 2020/12/18

Y1 - 2020/12/18

N2 - 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).

AB - 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).

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

Abstract

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