Antibiotic antimycotic solution

Human DPCs (ScienCell Research Laboratories, Carlsbad, CA, USA) were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Gibco), 10 ng/mL basic fibroblast growth factor (Peprotech, London, UK) and antibiotic/antimycotic solution (Gibco). Primary human LECs and blood vascular endothelial cells (BECs) isolated from foreskin were cultured as described previously [17 (link)]. LECs were incubated on 10 μg/mL fibronectin-coated dishes in endothelial cell basal medium (EBM; Lonza, Walkersville, MD, USA) containing 20% FBS, 2 mM l-glutamine (Gibco), antibiotic-antimycotic solution, 10 μg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA) and 25 μg/mL cAMP (Sigma-Aldrich). BECs were incubated on 10 μg/mL fibronectin-coated dishes in EBM supplemented with 20% FBS, 2 mM l-glutamine, antibiotic-antimycotic solution and 0.4% endothelial cell growth supplement (ECGS; PromoCell, Heidelberg, Germany). Cells were incubated at 37 °C and 5% CO₂ in a humidified incubator.

The methods used for culturing human DPCs have been described previously [31 (link), 32 (link)]. DPCs (ScienCell Research Laboratories, Carlsbad, CA, USA) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Gibco), 10 ng/mL basic fibroblast growth factor (Peprotech, London, UK) and antibiotic/antimycotic solution (Gibco). Primary human LECs and BECs isolated from foreskin were cultured as described previously [33 (link)]. LECs were cultured on 10 μg/mL fibronectin (Millipore, Billerica, MA, USA)-coated dishes in endothelial cell basal medium (EBM; Lonza, Walkersville, MD, USA), supplemented with 20% FBS, 2 mM l-glutamine (Gibco), antibiotic-antimycotic solution, 10 μg/mL hydrocortisone (Sigma-Aldrich) and 25 μg/mL cAMP (Sigma-Aldrich). BECs were cultured on 10 μg/mL fibronectin-coated dishes in EBM supplemented with 20% FBS, 2 mM l-glutamine, 1x antibiotic-antimycotic solution and 0.4% endothelial cell growth supplement (ECGS; PromoCell). Human dermal fibroblasts (DFs) were cultured in DMEM supplemented with 10% FBS and antibiotic/ antimycotic solution. Cells were incubated at 37°C in a 5% CO₂ incubator.