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The medium was composed of AdDMEM/F12 (Invitrogen) supplemented with 1% HEPES, 1% GlutaMAX (ThermoFisher), 1% penicillin/streptomycin (ThermoFisher), 1 B27 without retinoic acid (Gibco), 1.25 mM N -acetylcysteine (Sigma), 10 nM gastrin (Sigma) and the following growth factors: 50 ng ml 1 hEGF (Peprotech), 15% RSPO1 conditioned medium (home-made), 100 ng ml 1 FGF10 (Peprotech), 100 ng ml 1 FGF7 (Peprotech), 50 ng ml 1 HGF (Peprotech), 10 mM nicotinamide (Sigma, for EM1 medium only), 2 M A83-01 (Tocris), 3 M CHIR99021 (Tocris), 10 M Y-27632 (Tocris), 0.5 nM Wnt surrogate Fc fusion protein as in ref

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doi: 10.1021/bi990438f