Hematopoietic Differentiation and Production of Mature Myeloid Cells From Human Pluripotent Stem Cells

Nat Protoc. 2011 Mar;6(3):296-313. doi: 10.1038/nprot.2010.184. Epub 2011 Feb 17.

Abstract

In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin(-)CD34(+)CD43(+)CD45(+) multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5-19 d of culture with cytokines, depending on the cell type.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Differentiation*
  • Coculture Techniques / methods
  • Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Hematopoiesis / physiology
  • Humans
  • Myeloid Progenitor Cells / cytology
  • Myeloid Progenitor Cells / metabolism*
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*
  • Stromal Cells / cytology
  • Stromal Cells / metabolism*
  • Time Factors

Substances

  • Granulocyte-Macrophage Colony-Stimulating Factor