Fibronectin coated wells

For lineage depleted bone marrow cell preparation, linage marker positive cells were depleted using the biotin based selection kit (cat# 19856, STEMCELL Technologies, Vancouver, BC) according to the manufacturer’s instructions. Lin-BM cells were seeded in fibronectin-coated wells (Corning Inc, Corning, NY). To induce erythropoiesis, Lin- BM cells were cultured as described^{29 (link)}. On the first day, Lin- BM cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM medium) supplemented with 15% FBS, 1% detoxified bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO), 200 µg/mL holo-transferrin (Sigma-Aldrich, St. Louis, MO), 10 µg/mL recombinant human insulin (Sigma-Aldrich, St. Louis, MO), 2 mM L-glutamine, 10⁻⁴ M β-mercaptoethanol, 50 units/ml penicillin G, 50 µg/ml streptomycin (Thermo Fisher Scientific, Waltham, MA) and 2 U/mL Epo (STEMCELL Technologies, Vancouver, BC). The medium was replaced with IMDM with 20% FBS, 2 mM L-glutamine and 10⁻⁴ M β-mercaptoethanol for erythroid differentiation for the second and third day.

For lineage depleted bone marrow cell preparation, linage marker positive cells were depleted using the biotin based selection kit (cat# 19856, STEMCELL Technologies, Vancouver, BC) according to the manufacturer's instructions. Lin- BM cells were seeded in fibronectin-coated wells (Corning Inc, Corning, NY). To induce erythropoiesis, Lin- BM cells were cultured as described [24 (link)].