Additional file 1 of Effects of hydroxyapatite-coated nonwoven polyethylene/polypropylene fabric on non-mesodermal lineage-specific differentiation of human adipose-derived stem cells

Additional file 1. A schema of the experimental design. The methodology was as described elsewhere [27, 28]. ASCs were cultured until P3, assessed for cell viability, proliferation, MSC phenotype and multilineage potential. Then, the P3 ASCs were seeded at a similar density of 1 × 105 cells/mL in 1 mL of ADSC-GM medium on to the 2D and 3D platforms and cultured overnight. After 24 h, medium was replaced with ADSC-GM supplemented with 1% ITS + Premix and 10 μmol/L of 5‐Aza or ADSC-GM supplemented with 1% ITS + Premix only for the control experiments and cultured for another 24-h period (Day 0–1). Thereafter, cultures were washed 3X with PBS and maintained in fresh ITS + Premix-supplemented media every 3–4 days for 1–5 weeks. Cells were harvested weekly for cardiac phenotype monitoring by phase contrast microscopy and indirect immunocytochemical analysis (IFA).