Sequential analysis of alpha- and beta-globin gene expression during erythropoietic differentiation from primate embryonic stem cells

Stem Cells. 2006 Dec;24(12):2627-36. doi: 10.1634/stemcells.2006-0199. Epub 2006 Aug 3.


The temporal pattern of embryonic, fetal, and adult globin expression in the alpha (zeta --> alpha) and beta (epsilon --> gamma and gamma --> beta) clusters were quantitatively analyzed at the transcriptional and translational levels in erythrocytes induced from primate embryonic stem cells in vitro. When vascular endothelial growth factor receptor-2(high) CD34(+) cells were harvested and reseeded onto OP9 stromal cells, two-wave erythropoiesis occurred sequentially. Immunostaining and real-time reverse transcription-polymerase chain reaction analyses of floating mature erythrocytes revealed that globin switches occurred in parallel with the erythropoietic transition. Colony-forming assays showed replacement of primitive clonogenic progenitor cells with definitive cells during culturing. A decline in embryonic zeta- and epsilon-globin expression at the translational level occurred in individual definitive erythroid progenitors. Expression of beta-globin in individual definitive erythroid progenitors was upregulated in the presence of OP9 stromal cells. Thus, this system reproduces early hematopoietic development in vitro and can serve as a model for analyzing the mechanisms of the globin switch in humans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD34 / immunology
  • Coculture Techniques
  • Embryonic Stem Cells / cytology*
  • Erythrocytes / cytology
  • Erythropoiesis / genetics*
  • Flow Cytometry
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental*
  • Globins / genetics*
  • Humans
  • Macaca fascicularis / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stromal Cells / cytology
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism


  • Antigens, CD34
  • RNA, Messenger
  • Globins
  • Vascular Endothelial Growth Factor Receptor-2