Basic fibroblast growth factor bfgf

293T/17 cells (ATCC, Manassas, VA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; GIBCO BRL, Grand Island, NY) supplemented with 10% (v/v) Fetal Bovine Serum (FBS; GIBCO) and 1% (v/v) Antibiotic-Antimycotic (Anti-Anti; GIBCO). Human Hair Follicle derived Mesenchymal Stem Cells (hHF-MSCs) from a 73 year old male donor were isolated and characterized for differentiation potential as described previously [16 (link), 23 (link)] and human Bone Marrow derived Mesenchymal Stem Cells (hBM-MSCs, 29 year old male; Stem Cell Technologies, Vancouver, Canada) were cultured in growth medium (GM): DMEM supplemented with 10% (v/v) Mesenchymal Stem Cell qualified Fetal Bovine Serum (MSC-FBS; GIBCO), 1% (v/v) Anti-Anti and 1 ng/ml basic Fibroblast Growth Factor (bFGF; Biolegend, San Diego, CA). Cells were induced to myogenic differentiation using myogenic differentiation medium (DM): DMEM supplemented with 10% (v/v) MSC-FBS and 1% (v/v) Anti-Anti + 10 ng/ml TGF-β1 (Biolegend) + 30 μg/ml Heparin (APP Pharmaceuticals, LLC, Schaumburg, IL).

Human adipose-derived stromal cells (ASCs) were purchased from Lonza (Basel, Switzerland) and breast cancer cell lines MDA-MB-231 and MCF-7 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cells were expanded and cultured in growth medium (Dulbecco’s Modified Eagle’s Medium/Ham’s F-12 (DMEM/F12) w/o phenol red (Life Technologies, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS; Life Technologies) and 1% penicillin/streptomycin (100 U/mL penicillin, 0.1 mg/mL streptomycin; Life Technologies) at 37 °C and 5% CO₂. For the expansion of ASCs, 3 ng/mL basic fibroblast growth factor (bFGF; BioLegend, London, UK) was added to the growth medium. The medium was exchanged every other day. At 80–85% confluence, cells were passaged using 0.25% trypsin-EDTA solution (Life Technologies). ASCs at passage 6 were used for subsequent experiments.