Journal: BMC Cell Biology

Article Title: Generation of renewable mouse intestinal epithelial cell monolayers and organoids for functional analyses

doi: 10.1186/s12860-018-0165-0

Figure Lengend Snippet: Mouse small intestinal epithelial cells divide and maintain morphology with passage and express epithelial cell markers when cultured under modified conditional reprogramming conditions. a Representative phase contrast images of IEC monolayer colonies 1 day post-plating and 4 days after post-plating (passage 12). Dashed circle indicates colonies. Images were taken at 10× magnification. Scale bar: 100 μm. b Representative image of colony with higher magnification (20×) c Representative flow cytometry plot of EpCAM expression in IEC monolayers. Data represents IEC monolayers from 3 individual mice and diverse passages. d Representative phase contrast images of IEC monolayers colonies at early, middle and later passages. e Representative fluorescence microscopy images of IEC monolayers stained with tight junction markers E-cadherin and ZO-1

Article Snippet: E-cadherin (1:100; Cat# 3195; Cell Signaling Technology, Danvers, MA) and ZO-1 (1:25; Clone# R40.76; a kind gift from Dr. Alan Fanning; UNC-Capel Hill) antibodies were diluted in permeabilization buffer (5% normal goat serum/0.5% Triton-X in PBS) and incubated on cells overnight at 4 °C.

Techniques: Cell Culture, Modification, Flow Cytometry, Cytometry, Expressing, Mouse Assay, Fluorescence, Microscopy, Staining