Immortalization of erythroblasts by c-MYC and BCL-XL enables large-scale erythrocyte production from human pluripotent stem cells

Stem Cell Reports. 2013 Dec 5;1(6):499-508. doi: 10.1016/j.stemcr.2013.10.010. eCollection 2013.

Abstract

The lack of knowledge about the mechanism of erythrocyte biogenesis through self-replication makes the in vitro generation of large quantities of cells difficult. We show that transduction of c-MYC and BCL-XL into multipotent hematopoietic progenitor cells derived from pluripotent stem cells and gene overexpression enable sustained exponential self-replication of glycophorin A(+) erythroblasts, which we term immortalized erythrocyte progenitor cells (imERYPCs). In an inducible expression system, turning off the overexpression of c-MYC and BCL-XL enabled imERYPCs to mature with chromatin condensation and reduced cell size, hemoglobin synthesis, downregulation of GCN5, upregulation of GATA1, and endogenous BCL-XL and RAF1, all of which appeared to recapitulate normal erythropoiesis. imERYPCs mostly displayed fetal-type hemoglobin and normal oxygen dissociation in vitro and circulation in immunodeficient mice following transfusion. Using critical factors to induce imERYPCs provides a model of erythrocyte biogenesis that could potentially contribute to a stable supply of erythrocytes for donor-independent transfusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Differentiation / genetics
  • Cell Size
  • Embryonic Stem Cells
  • Erythroblasts / cytology
  • Erythroblasts / metabolism*
  • Erythropoiesis / genetics*
  • GATA1 Transcription Factor / genetics
  • GATA1 Transcription Factor / metabolism
  • Gene Expression Regulation
  • Genes, myc*
  • Hemoglobins / biosynthesis
  • Humans
  • Mice, Inbred NOD
  • Mice, SCID
  • Oxygen / metabolism
  • Pluripotent Stem Cells
  • Proto-Oncogene Proteins c-myc / genetics*
  • Proto-Oncogene Proteins c-myc / metabolism
  • Proto-Oncogene Proteins c-raf / genetics
  • Proto-Oncogene Proteins c-raf / metabolism
  • Transduction, Genetic
  • bcl-X Protein / genetics*
  • bcl-X Protein / metabolism
  • p300-CBP Transcription Factors / genetics
  • p300-CBP Transcription Factors / metabolism

Substances

  • GATA1 Transcription Factor
  • GATA1 protein, human
  • Hemoglobins
  • Proto-Oncogene Proteins c-myc
  • bcl-X Protein
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor
  • Proto-Oncogene Proteins c-raf
  • Oxygen