Erythroid Differentiation of hPSC-derived Progenitors

Erythroid differentiation was performed using the StemSpan Erythroid Expansion Supplement (STEMCELL Technologies, 02692) following the manufacturer’s recommendations. In brief, day 10 hPSC-derived HLF+ HOXA+ hematopoietic progenitors were collected, counted, and seeded in StemSpan SFEM II (STEMCELL Technologies, 09655) supplemented with the StemSpan Erythroid Expansion Supplement at a density of 1x10⁴-1x10⁵ cells/mL. On day 3, cells were supplemented with an equal volume of complete media. On day 7 and 10, cells were harvested and replated in complete media at a density of 1x10⁵ cells/mL. On day 14, cells were harvested and processed for flow cytometry.
High-performance liquid chromatography (HPLC) to detect hemoglobin tetramers was performed as previously described.²⁰⁶ (link)