fig1

Figure 1. GuMI can coculture F. prausnitzii A2-165 with colonic epithelium from multiple donors for up to 4 days. (A) The general workflow of the GuMI experiment includes monolayer seeding, hardware preparation, GuMI assembly, bacterial injection, and sampling; (B) TEER value of monolayers at 48 hours after bacterial injection under static condition (Static), GuMI-FP or GuMI-NB. The monolayers were derived from an uninflamed/unaffected region in the transverse colon from donor H462, a 15-year-old male with CD. *P < 0.05, ***P < 0.001, one-way ANOVA was performed with Tukey’s multiple comparisons test; (C) Brightfield images of monolayers at the end of the GuMI experiment in Static, GuMI-NB, and GuMI-FP; (D) Growth of F. prausnitzii after 48 h of coculture in GuMI. *P < 0.05 two-tailed unpaired t-test; (E) Brightfield images of monolayers. Coculture of F. prausnitzii with two donors, HC2978 and HC465 (ulcerative colitis patient), was maintained for four days. Dendritic cells and macrophages were included underneath the monolayers; (F) TEER values of monolayers in GuMI. P = 0.55, two-tailed unpaired t-test; (G) Bacterial density in the apical compartment of GuMI after four days of coculture compared to the inoculum (0d). Note that the same inoculum (0d) applies to HC2978 and HC465. *P < 0.05, **P < 0.01, two-tailed unpaired t-test. TEER: Transepithelial electric resistance; GuMI-FP: GuMI with F. prausnitzii; GuMI-NB: GuMI without bacteria.