Hi,

I am currently working on the establishment of intestinal monolayer cultures from mouse colonic organoid cultures on 3.0 µm pore size filters.

I have already established a protocol to culture them on 0.4 µm pore size filters. The state of confluency is measured by transepithelial resistance measurements (TEER).

Unfortunately, the cells won't grow on the 3.0µm filters. I have coated the filters either with 1:30 Matrigel dilution (as for the 0.4 µm) or with a rehydrated Collagen I overlaid with a lower dilution of Collagen (both from published articles). Nevertheless I don't see a build-up in resistance, even though some cell patchy are detectable with IHC.

Is anyone maybe also working with mouse intestinal organoids and the generation of monolayers on the 3.0µm filters and has some suggestions what could be the problem?