Hematopoietic Differentiation of iPSCs

iPSCs were obtained from the iPSC repository of the DIAN project, directed by Dr. Karch (Washington University, St. Louis, MO, USA) [49 (link)]. Donor line information is detailed in Table 1. iPSC lines were thawed into Matrigel-coated 6-well plates and maintained by feeding every other day with Stemflex supplemented media (Thermo Fisher Scientific, Waltham, MA, US) at 37 °C, 5% CO₂. When 70–80% confluence was reached, iPSCs were passaged using ReLeSR dissociation reagent (#05872, STEMCELL Technologies, Vancouver, Canada). Hematopoietic stem cells were differentiated from iPSCs using the STEMdiff Hematopoietic kit (#05310 STEMCELL Technologies, Vancouver, Canada,) according to a previously published protocol [41 (link)]. When cultured for a minimum of 10 days, HPCs were collected and, if viability was >75%, frozen in Bambanker media (Wako Chemicals, Osakan, Japan).