Differentiation of reprogrammed α-thalassemia fibroblasts. (A) H&E staining demonstrating that iPSCs derived from α-thalassemia fibroblasts with transgene-free method can form teratoma in NSG mice, and generate cells from the 3 germ layers (bar = 50 μm). (B) HPLC analysis of globin expression and morphology of the erythroblasts obtained after differentiation of α⁰-thal-iPSCs and control H1 hESCs. (Top panels) Basophilic erythroblasts obtained after 14 days of coculture on FH-B-hTERT and 14 days of liquid culture. (Bottom panels) Orthochromatic erythroblasts obtained after an additional 10 days of liquid culture. (Left panels) HPLC profiles and Giemsa staining of cells obtained after differentiation of α⁰-thal-iPSCs. (Right panels) Same as left panels but for control H1 hESCs. α⁰-thal-iPSCs do not express any α-globin chains. Zeta-globin chains are silenced between the 14th and the 24th day of liquid culture.