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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‐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 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.

Sequential Analysis of α‐ and β‐Globin Gene Expression During Erythropoietic Differentiation from Primate Embryonic Stem Cells