Figure 2.
A, Gene correction of the sickle mutation in β‐globin gene in sickle cell disease (SCD)‐induced pluripotent stem cells (iPSCs), followed by our optimized serum‐free iPSC‐derived sac (iPS‐sac) generation and serum‐free erythroid differentiation. B, Nucleotide‐specific chromatogram peaks for monoallelic correction (mCOR, left panel) and biallelic correction (bCOR, right panel), analyzed by Sanger sequencing. C, Chromosomes with a Giemsa‐banding stain in a biallelic corrected iPSC line. D, Yields of iPS‐sac‐derived hematopoietic‐like spherical cells per iPSC at day 19 (left panel) and erythroid cells per iPSC after differentiation of the spherical cells (right panel). E, Red colored pellets of iPS‐sac‐derived erythroid cells from non‐corrected SCD iPSCs as well as gene‐corrected iPSCs (mCOR and bCOR). F, Percentages of cell surface markers in iPS‐sac‐derived spherical cells from SCD‐iPSCs, mCOR‐iPSCs, and bCOR‐iPSCs. G, Normal β‐globin and βS‐globin production at the protein level, analyzed by RP‐HPLC. Data reported as mean ± SD. Statistical analysis was performed by Tukey's honestly significant difference test (*P < .05 and **P < .01). SCD‐iPS, n = 4; mCOR‐iPS, n = 4; bCOR‐iPS, n = 4 for all the analysis