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To isolate intestinal organoid, following euthanasia of 810-week-old male mice, crypts were isolated from the ileum fragments by EDTA incubation and plated in Matrigel using advanced DMEM/F12 containing 100 U/mL penicillin-streptomycin, 10 mM HEPES, 2 mM GlutaMAX (35050061), supplements N2 (17502048) and B27 (17504044) from ThermoFisher, 50 ng/mL murine EGF (315-09, Peprotech), 1 mmol/L N-acetylcysteine (A9165, Sigma), 100 ng/mL murine Noggin (250-38, Peprotech), and 1000 ng/mL human R-spondin-1 (120-38-20, Peprotech)
JensterleMFerjanSLeaiLSoanAGoriarKZaletelKet al
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doi: 10.1016/S2213-8587(19)30423-1 89 BasalayMVDavidsonSMYellonDM
The 10 mg dose represents the threshold where many patients begin experiencing maximal weight reduction trajectories
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