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

STAR Protoc. 2020 Oct 16;1(3):100130. doi: 10.1016/j.xpro.2020.100130. eCollection 2020 Dec 18.

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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