Primary hematopoietic stem and progenitor cells (HSPCs) were obtained as described in (28 (link)). Briefly, cKit magnetic beads (Miltenyi Biotec) were used to enrich HSPCs from mouse bone marrow and cultured in IMDM (Gibco) supplemented with 15% FBS (Gibco), 10 ng/ml mIL-6, 10 ng/ml mIL-3, 50 ng/ml SCF, 20 ng/ml thrombopoietin (PeproTech) and 10 ng/ml Flt3 ligand (PeproTech).
The E14 and K562 cell lines were obtained from the experimental pathology cell bank in State Key Laboratory of Experimental Hematology. The K562 cells were cultured in RPMI-1640 (Gibco) supplemented with 10% FBS. E14 cells were maintained in 0.2% gelatin (Sigma)-coated plates in 2i medium, which consisted of DMEM/F12 supplemented with Neurobasal medium, serum-free N2B27 medium supplemented with 10 μM MEK inhibitor PD0325901 and 30 μM GSK3 inhibitor CHIR99021 (both from Selleckchem), 2% KnockOut™ serum replacement (Gibco), 0.002% BSA (Gibco), 1 mM MTG (Gibco), 1000 U/ml LIF (Millipore), 0.1 mM non-essential amino acid and 2 mM GlutaMAX Supplement (Gibco).
The E14 and K562 cell lines were obtained from the experimental pathology cell bank in State Key Laboratory of Experimental Hematology. The K562 cells were cultured in RPMI-1640 (Gibco) supplemented with 10% FBS. E14 cells were maintained in 0.2% gelatin (Sigma)-coated plates in 2i medium, which consisted of DMEM/F12 supplemented with Neurobasal medium, serum-free N2B27 medium supplemented with 10 μM MEK inhibitor PD0325901 and 30 μM GSK3 inhibitor CHIR99021 (both from Selleckchem), 2% KnockOut™ serum replacement (Gibco), 0.002% BSA (Gibco), 1 mM MTG (Gibco), 1000 U/ml LIF (Millipore), 0.1 mM non-essential amino acid and 2 mM GlutaMAX Supplement (Gibco).
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