Peripheral blood human plerixafor or granulocyte colony-stimulating factor (G-CSF)-mobilized adult HSPCs or cord blood HSPCs were obtained from HDs. Peripheral blood non-mobilized or plerixafor-mobilized human adult HSPCs were obtained from SCD patients. SCD and HD samples eligible for research purposes were obtained from the Necker-Enfants malades Hospital (Paris, France) except plerixafor-mobilized HD cells that were purchased from Caltag and Hemacare. Written informed consent was obtained from all the subjects. All experiments were performed in accordance with the Declaration of Helsinki. The study was approved by the regional investigational review board (reference: DC 2022-5364, CPP Ile-de-France II "Hôpital Necker-Enfants malades"). HSPCs were purified by immunomagnetic selection after immunostaining using the CD34 MicroBead Kit (Miltenyi Biotec). HSPCs were thawed and cultured at a concentration of 5x10 5 cells/ml in the "HSPC medium"
containing StemSpan (STEMCELL Technologies) supplemented with penicillin/streptomycin (Gibco), 250 nM StemRegenin1 (STEMCELL Technologies), 20 nM UM171 (STEMCELL Technologies), and the following recombinant human cytokines (PeproTech): human stem cell factor (SCF; 300 ng/ml), Flt-3L (300 ng/ml), thrombopoietin (TPO; 100 ng/ml), and interleukin-3 (IL-3; 60 ng/ml).
containing StemSpan (STEMCELL Technologies) supplemented with penicillin/streptomycin (Gibco), 250 nM StemRegenin1 (STEMCELL Technologies), 20 nM UM171 (STEMCELL Technologies), and the following recombinant human cytokines (PeproTech): human stem cell factor (SCF; 300 ng/ml), Flt-3L (300 ng/ml), thrombopoietin (TPO; 100 ng/ml), and interleukin-3 (IL-3; 60 ng/ml).
