Sequential Analysis of α- and β-Globin Gene Expression During Erythropoietic Differentiation from Primate Embryonic Stem Cells Academic Article uri icon

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abstract

  • 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.

authors

  • Umeda, Katsutsugu
  • Heike, Toshio
  • Nakata-Hizume, Mami
  • Niwa, Akira
  • Arai, Masato
  • Shinoda, Gen
  • Ma, Feng
  • Suemori, Hirofumi
  • Luo, Hong Yuan
  • Chui, David Hing-kwei
  • Torii, Ryuzo
  • Shibuya, Masabumi
  • Nakatsuji, Norio
  • Nakahata, Tatsutoshi

publication date

  • December 2006

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