Application of small molecule CHIR99021 leads to the loss of hemangioblast progenitor and increased hematopoiesis of human pluripotent stem cells

Exp Hematol. 2018 Sep:65:38-48.e1. doi: 10.1016/j.exphem.2018.05.007. Epub 2018 Jun 5.

Abstract

Improving our understanding of the intricacies of hematopoietic specification of induced or embryonic human pluripotent stem cells is beneficial for many areas of research and translational medicine. Currently, it is not clear whether, during human pluripotent stem cells hematopoietic differentiation in vitro, the maturation of definitive progenitors proceeds through a primitive progenitor (hemangioblast) intermediate or if it develops independently. The objective of this study was to investigate the early stages of hematopoietic specification of pluripotent stem cells in vitro. By implementing an adherent culture, serum-free differentiation system that utilizes a small molecule, CHIR99021, to induce human pluripotent stem cells toward various hematopoietic lineages, we established that, compared with the OP9 coculture hematopoietic induction system, the application of CHIR99021 alters the early steps of hematopoiesis such as hemangioblasts, angiogenic hematopoietic progenitors, and hemogenic endothelium. Importantly, it is associated with the loss of hemangioblast progenitors, loss of CD43+ (primitive hematopoietic marker) expression, and predominant development of blast-forming unit erythroid colonies in semisolid medium. These data support the hypothesis that the divergence of primitive and definitive programs during human pluripotent stem cells differentiation precedes the hemangioblast stage. Furthermore, we have shown that the inhibition of primitive hematopoiesis is associated with an increase in hematopoietic potential, which is a fruitful finding due to the growing need for lymphoid and myeloid cells in translational applications.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Differentiation / drug effects*
  • Cell Line
  • Cell Lineage
  • Erythroid Cells / cytology
  • Erythroid Cells / drug effects
  • Hemangioblasts / cytology*
  • Hematopoietic Stem Cells / cytology*
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / drug effects*
  • Humans
  • Microscopy, Confocal
  • Pluripotent Stem Cells / cytology*
  • Pyridines / pharmacology*
  • Pyrimidines / pharmacology*
  • Real-Time Polymerase Chain Reaction

Substances

  • Chir 99021
  • Pyridines
  • Pyrimidines