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Crypt-laden Matrigel was dispensed onto 48-well culture plates, and following solidification of Matrigel droplets, the crypts were overlaid with the following medium: Advanced Dulbeccos Modified Eagles Medium/F12 (Thermon Fisher Scientific) supplemented with penicillin/streptomycin, 10 mM HEPES, 2 mM GlutaMAX, 1 B-27 Supplement (Thermo Fisher Scientific), 10 nM gastrin I (Sigma-Aldrich), 1 mM N-acetylcysteine (Sigma-Aldrich), and the niche factors of 100 ng/mL recombinant mouse Noggin (PeproTech), 50 ng/mL recombinant mouse EGF (Invitrogen), 10% R-spondin1 conditioned medium, 500 nM A83-01 (Tocris), 100 ng/mL human Wnt-3 (R&D system)

Eat more sulfur-rich foods like garlic and cruciferous vegetables and make sure you get enough selenium

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Olson K, Dover JS, Arndt KA