Figure 1.

Schematic of ALI + VIP culture of colonic cells followed by mucus collection. (a) Cells (green) are seeded on a microporous insert (gray) with expansion medium (orange) in both the apical and basal reservoirs. (b) Cells are grown into a confluent monolayer over the course of 3–5 days. (c) After confluence is achieved, both the apical and basal media are aspirated. Differentiation medium with VIP (red) is placed in the basal reservoir underneath the microporous insert. No medium is added to the apical reservoir. A hydrated continuous mucus layer (yellow) forms on the surface of the epithelial cells. (d) After 2–7 days of culture, the mucus layer is collected.