Antibiotic antimyotic solution

Lin⁻cKit⁺ HSPCs were cultured in StemSpan® serum-free expansion medium (SFEM) containing 100 ng/mL recombinant mouse SCF, 6 ng/mL recombinant mouse IL3 and 10 ng/mL recombinant human IL-6 (all from Stem Cell Technologies, Vancouver, BC)
ECs were cultured on 1% gelatin coated plates in complete EC medium which is consisted of advanced DMEM/F12 (ThermoFisher, Waltham, MA) medium containing 20% fetal bovine serum, 50μg/ml endothelial cell growth supplement (Alfa Aesar, Ward Hill, MA), 1% Antibiotic-antimyotic solution (Cat. 15240-062, ThermoFisher), 10mM HEPES buffer (ThermoFisher), 5μM SB431542 small molecule (R&D, Minneapolis, MN), 50μg/ml Heparin (Sigma), 1% Glutamax 100× solution (ThermoFisher), 1% non-essential amino acid (ThermoFisher), recombinant mouse VEGF 10ng/ml (PeproTech, Rocky Hill, NJ; add fresh when changing medium) and recombinant human FGF2 20ng/ml (PeproTech; add fresh when changing medium). Cells were cultured at 37°C in a humidified 5% CO₂ atmosphere with medium change every 2–3 days until they reach 70–80% confluence. 0.05 % trypsin-EDTA solution was used for cell passaging. EC-conditioned media (ECCM) was collected when cell density reach 50–60% confluency.

Lin^-cKit⁺ HSPCs were cultured in StemSpan® serum-free expansion medium (SFEM) containing 100 ng/mL recombinant mouse SCF, 6 ng/mL recombinant mouse IL3 and 10 ng/mL recombinant human IL-6 (all from Stem Cell Technologies, Vancouver, BC)
ECs were cultured on 2% gelatin coated plates in complete EC medium which is consisted of advanced DMEM/F12 (ThermoFisher, Waltham, MA) medium containing 20% fetal bovine serum, 50μg/ml endothelial cell growth supplement (Alfa Aesar, Ward Hill, MA), 1% Antibiotic-antimyotic solution (Cat. 15240–062, ThermoFisher), 10mM HEPES buffer (ThermoFisher), 5μM SB431542 small molecule (R&D, Minneapolis, MN), 50μg/ml Heparin (Sigma), 1% Glutamax 100x solution (ThermoFisher), 1% non-essential amino acid (ThermoFisher), recombinant mouse VEGF 10ng/ml (PeproTech,Rocky Hill, NJ; add fresh when changing medium) and recombinant human FGF2 20ng/ml (PeproTech; add fresh when changing medium). Cells were cultured at 37C in a humidified 5% CO₂ atmosphere with medium change every 3 days until they reach 70–80% confluence. 0.05 % trypsin-EDTA solution was used for cell passaging. EC-conditioned media (ECCM) was collected when cell density reach 50–60% confluency.