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Add the melted butter, pumpkin puree, egg and vanilla - stir until well mixed

Materials and methods Materials Nunc EasYFlask 225 cm 2 (T225) cell culture flasks, Corning Costar six-well clear tissue culture-treated flat bottom plates, Falcon 50 mL conical centrifuge tubes, Basix microcentrifuge tubes, Invitrogen Countess cell counting chamber slides, Nunc 25 mL serological pipettes, Greiner Bio-One CELLSTAR 24-well cell culture plates, Corning 96-well solid white non-sterile microplates, 9 and 5 glass Pasteur pipets, Gibco trypsinEDTA (0.25%) phenol red, Gibco DMEM/F-12 with l-glutamine and phenol red, Corning penicillinstreptomycin (pen-strep) solution (10,000 IU penicillin, 10,000 g/mL streptomycin), and Corning regular fetal bovine serum (FBS) were purchased from Fisher Scientific (Pittsburgh, PA, USA)