D glucose g7528

Dose-response experiments using organoids were performed using additional D-(-)-fructose (F0127; Sigma-Aldrich), D-(+)-glucose (G7528; Sigma-Aldrich), sucrose (S0389; Sigma-Aldrich) at 200, 100, 50, 25, 12.5, 3.1, and 0.8 mmol/L concentrations (equaling 225, 125, 75, 50, 37.5, 28.1, and 25.8 mmol/L of glucose, respectively, adding in the base amount in DMEM). Briefly, normal colon organoids were dissociated to near-single cells and plated onto a 24-well plate, with 20-μL domes per well. A total of 500 μL culture medium further supplemented with different concentrations of D-(-)-fructose (F0127; Sigma-Aldrich), D-(+)-glucose (G7528; Sigma-Aldrich), or sucrose (S0389; Sigma-Aldrich) was added to each well at the time of seeding. Media with the supplemented sugars was refreshed every 2 days, and growth was followed up to day 12. Alternatively, normal colon organoids were dissociated to near-single cells and plated onto a 24-well plate, with 20-μL domes per well. A total of 500 μL standard culture medium was added and organoids were allowed to grow to day 5. Media then was changed to media supplemented with additional glucose, fructose, or sucrose at varying concentrations. Growth was followed up for the next 48 hours.

p-CA, LPS from Escherichia coli (L2637), HEPES (H4034), L-glutamine (G-8540) and D-glucose (G-7528) were purchased from Sigma (St. Louis, MO, USA). Minimum essential medium (MEM, LM 007-01), Hank's balanced salt solution (HBSS, LB 003-01) and horse serum (S 104-01) were purchased from JBI (Daegu, South Korea). Penicillin and streptomycin were obtained from Gibco BRL (LS 202-02, USA).