Figure 2 | Scientific Reports

Figure 2

From: Self-organized intestinal epithelial monolayers in crypt and villus-like domains show effective barrier function

Figure 2

Stem cells self-assemble in crypt-like domains and originate self-sustainable epithelial monolayers. (A) Live-imaging sequence of overlapped bright field and GFP signal corresponding to 60 hours after seeding organoid-derived crypts and (B) 96 hours after seeding organoid-derived single cells on a thin film of Matrigel. The corresponding time for each snapshot is shown in each panel. White arrow heads indicate Lgr5-GFP+ cells. Scale bars: 50 µm (A, upper row) and 100 µm (A, lower row, and B). (C) Immunofluorescence of EdU, Ki67, and ZO-1 of an EdU pulse-chase experiment. Representative images of 0 h, 24 h, 48 h, 72 h, and 96 h after EdU chase of either crypt-pieces (upper panels) or single cell (lower panels) derived cultures. Scale bars: 50 µm. (D) Immunofluorescence for Ki67 and F-actin of epithelial monolayers from passage 0 to passage 3 (P0, P1, P2, P3). Epithelial monolayers were passed by enzymatical digestion after every 6 to 8 days in culture. Scale bars: 100 µm.

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