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The samples were cultured in pancreatic organoid feeding medium, which consisted of advanced DMEM/F12 supplemented with 10 mM HEPES (Thermo Fisher Scientific, 15630056), 1 GlutaMAX (Thermo Fisher Scientific, 35050087), penicillinstreptomycin, 100 ng ml 1 animal-free recombinant human FGF10 (Thermo Fisher Scientific, 500-P151G-50UG), 50 ng ml 1 animal-free recombinant human EGF (Thermo Fisher Scientific, AF-100-15-1MG), 100 ng ml 1 recombinant human noggin (Bio-Techne, 6057-NG), WNT3A-conditioned medium (30% v/v), RSPO1-conditioned medium (10% v/v), 10 nM human gastrin 1 (Sigma-Aldrich, SCP0152), 10 mM nicotinamide (Sigma-Aldrich, N0636), 1.25 mM N -acetylcysteine (Sigma-Aldrich, A9165), 1 B27 (Thermo Fisher Scientific, 17504001), 500 nM A83-01 (Tocris, 2939/10) and 10.5 M Y27632 (Tocris, 1254/1)

After 90 min of incubation at 37C in an atmosphere of 5% CO 2 and 95% humidity, cells were harvested, washed with PBS and frozen in liquid nitrogen

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The most striking variations were observed through NADP-ME isozymic variations as resolved with NBT complex on non-denaturing gel (Fig