Directed differentiation of primitive and definitive hematopoietic progenitors from human pluripotent stem cells

Carissa Dege, Christopher M. Sturgeon

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


One of the major goals for regenerative medicine is the generation and maintenance of hematopoietic stem cells (HSCs) derived from human pluripotent stem cells (hPSCs). Until recently, efforts to differentiate hPSCs into HSCs have predominantly generated hematopoietic progenitors that lack HSC potential, and instead resemble yolk sac hematopoiesis. These resulting hematopoietic progenitors may have limited utility for in vitro disease modeling of various adult hematopoietic disorders, particularly those of the lymphoid lineages. However, we have recently described methods to generate erythro-myelo-lymphoid multilineage definitive hematopoietic progenitors from hPSCs using a stage-specific directed differentiation protocol, which we outline here. Through enzymatic dissociation of hPSCs on basement membrane matrix-coated plasticware, embryoid bodies (EBs) are formed. EBs are differentiated to mesoderm by recombinant BMP4, which is subsequently specified to the definitive hematopoietic program by the GSK3β inhibitor, CHIR99021. Alternatively, primitive hematopoiesis is specified by the PORCN inhibitor, IWP2. Hematopoiesis is further driven through the addition of recombinant VEGF and supportive hematopoietic cytokines. The resulting hematopoietic progenitors generated using this method have the potential to be used for disease and developmental modeling, in vitro.

Original languageEnglish
Article numbere55196
JournalJournal of Visualized Experiments
Issue number129
StatePublished - Nov 1 2017


  • Cell culture
  • Definitive hematopoiesis
  • Developmental biology
  • Embryoid bodies
  • Hemogenic endothelium
  • Human embryonic stem cells
  • Issue 129
  • Pluripotent stem cells
  • Primitive hematopoiesis
  • WNT signaling


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