Tpp tissue culture dishes

Bovine myoblasts and bovine adMSCs were sub-cultured on TPP® tissue culture dishes (Sigma Aldrich). The cell subculture was conducted using 1X phosphate buffered saline (PBS), 0.025% trypsin-EDTA, and the growth medium of each cell group. The cultured bovine myoblast (Passage 3) and adMSC (Passage 3) were seeded on the hydrogels. The seeding density of the myoblast and adMSC were 3 × 10⁴ cells/12 well molded scaffold and 2 × 10⁴ cells/12 well molded scaffold, respectively. The myoblasts were proliferated for 4 days, after which the medium was replaced with the myoblast differentiation medium, which is composed of HG-DMEM supplemented with 5% (v/v) horse serum and 1% (v/v) penicillin-streptomycin-glutamine (PS). Further, the adMSCs were proliferated for 3 days to achieve a cell confluency of approximately 90%, after which the medium was replaced with the adipose differentiation medium. The adipose differentiation medium was composed of LG-DMEM supplemented with 5% (v/v) FBS, 10 μM insulin from bovine pancreas, 1 μM dexamethasone, 10 μM ciglitizone, and 100 μM oleic acid. The medium was replaced every other day.