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- Title
Sequential Analysis of α- and β-Globin Gene Expression During Erythropoietic Differentiation from Primate Embryonic Stem Cells.
- Authors
Umeda, Katsutsugu; Heike, Toshio; Nakata-Hizume, Mami; Niwa, Akira; Arai, Masato; Shinoda, Gen; Feng Ma; Suemori, Hirofumi; Hong Yuan Luo; Chui, David H. K.; Ryuso Torii; Shibuya, Masabumi; Nakatsuji, Norio; Nakahata, Tatsutoshi
- Abstract
The temporal pattern of embryonic, fetal, and adult globin expression in the α (ζ →α) and β (ϵ → γ and γ → β) 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-2high 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 ζ- and ϵ-globin expression at the translational level occurred in individual definitive erythroid progenitors. Expression of β-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.
- Subjects
EMBRYONIC stem cells; GLOBIN genes; GENE expression; ERYTHROPOIESIS; VASCULAR endothelial growth factors; DNA polymerases; POLYMERASE chain reaction
- Publication
Stem Cells, 2006, Vol 24, Issue 12, p2627
- ISSN
1066-5099
- Publication type
Article
- DOI
10.1634/stemcells.2006-0199