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Spheroid assays A total of 1 10 4 cells were seeded in 24-well ultra-low attachment culture plates (Corning, NY, USA) in the presence of the specified inhibitor(s) in serum-free media supplemented with 20 ng/ml EGF (Gibco), 20 ng/ml bFGF (Gibco), 10 g/ml heparin (Sigma-Aldrich), 1:50 (v/v) B-27 supplement 50x (Gibco), 5 g/ml human insulin (Sigma-Aldrich) and 0.5 g/ml hydrocortisone (Sigma-Aldrich)

Comparison with Other Compounds BPC-157 and TB-500 differ from metabolic peptides such as Tirzepatide 40mg research peptide, as they are primarily explored for recovery and regeneration rather than metabolic signalling

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