Establishment of Epithelial Inflammatory Injury Model Using Intestinal Organoid Cultures

Figure 2

Proliferation ability assay of intestinal organoid with TNF-α treatment. (a) qRT-PCR analyses of proliferation marker Ki67 and ISC marker Lgr5 expression in organoids cultured under the indicated conditions ( well). values were determined by two-sided unpaired Student’s -test. Gapdh was used as a control to normalize the expression of target genes. ; ; . (b) Immunofluorescence staining of Ki67 (green) of intestinal organoids after 3 days of culture in groups TNF-α-10 (first column) and ENR (second column). Images of typical giant organoids (first row) and budding organoids (second row). Blue stain (DAPI) indicates the nucleus. Scale  μm. White arrows pointed to the Ki67⁺ cells, and red arrows pointed to typical buddings. (c) The ratio of the number of giant organoids and the total number of intestinal organoids (giant organoids plus budding organoids). DAPI nucleus labeling was used in morphological differences of organoid cultures to visualize the budding organoids and giant organoids. ; ; . The experiments (a, c) were independently repeated at least three times with similar results.

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