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. Author manuscript; available in PMC: 2019 Mar 12.
Published in final edited form as: Dev Cell. 2018 Mar 1;44(5):624–633.e4. doi: 10.1016/j.devcel.2018.01.024

Figure 1.

Figure 1

Enteroid monolayers establish proliferative focal patterning and contain differentiated epithelial cell types. (A) Bright field images of 2D intestinal sheet 7 days after seeding. Arrowheads mark putative crypt foci. Scale bar = 25μM. (B) 2D intestinal cultures treated with EdU for 2 hours, then fixed and stained for EdU and DAPI. EdU clusters and dense nuclei foci co-localize (merged), indicating that the dense nuclear regions are proliferative crypt foci. Scale bar = 25μM. (C) Confocal image of crypt foci showing proliferative cells (EdU, green) clustered with Paneth cells (UEA-1, red) surrounded by non-proliferative cells (DAPI, blue). Scale bar = 25μM. (D) Confocal image of enteroid monolayers stained for actin (green) and DNA (blue). Arrowheads mark apical actin bundles. Right and bottom panels in c.–d. indicate horizontal (H) and vertical (V) projections. Scale bar = 25μM. (E) Enteroid monolayer cultures from Lgr5eGFP-DTR mice showing Lgr5+ stem cells (green) juxtaposed with Paneth cells (lysozyme, red) and surrounded by proliferative TA cells (EdU, blue). Scale bar = 25μM. (F) Enteroid culture images co-stained for cell-fate marker (red: Ki-67 – proliferative; UEA-1 – Paneth; Muc2 – Goblet; villin – enterocyte; DCLK-1 – Tuft; ChgA – enteroendocrine; Olfm4 – stem), EdU (green) and DAPI (blue). Scale bar = 25μM. Crypts in B–F were seeded 7 days before staining and imaging. See also Figures S1, S2, and S3.