Fig. 4

Hematopoietic differentiation of iPSCs using the OP9 coculture system and erythroid liquid culture. a Schematic of hematopoietic differentiation protocol used in this study. b Morphological changes of the Eβ-iPSC2 cells and the corrected C46 cells during hematopoietic differentiation on day 6 of OP9 coculture (scale bar = 500 μm), and Wright’s staining on days 13, 23 and 29 of differentiation. c Quantitative RT-PCR analysis of erythroid-associated transcription factors at day 19 of differentiation (equivalent to day 13 of erythroid liquid culture) of the Eβ-iPSC2 cells and the corrected C46 cells as compared to peripheral blood CD34⁺ cell-derived erythroblasts at day 13 (PB). Data presented as mean ± SD of triplicate samples from a representative experiment. d Western blot analysis of alpha and beta hemoglobin expression of the Eβ-iPSC2 cells and the corrected C46 cells at day 30 of differentiation as compared to peripheral blood CD34⁺ cell-derived erythroblasts at day 24 of erythroid liquid culture. MEM minimal essential medium, IMDM Iscove modified Dulbecco medium, FBS fetal bovine serum, SCF stem cell factor, IL interleukin, EPO erythropoietin, Eβ-iPSC2 iPSC lines derived from a patient with HbE/β-thalassemia, Hb hemoglobin