Fig. 1: Characterization of crypt-like and villus-like human small intestinal organoids. | Cell Death & Disease

Fig. 1: Characterization of crypt-like and villus-like human small intestinal organoids.

From: Proteomics analysis of human intestinal organoids during hypoxia and reoxygenation as a model to study ischemia-reperfusion injury

Fig. 1

A Brightfield images representing different phenotypes of hSIOs cultured in complete growth medium (GM) for 12 days (upper panel, scale bar = 200 µm) and differentiated hSIOs grown in GM for 7 days followed by 5 days in differentiation medium (DM) (lower panel, scale bar = 100 µm). B mRNA expression of crypt cell markers OLFM4 (stem cells) and LYZ (Paneth cells) and villus cell markers I-FABP (absorptive enterocytes) and MUC2 (Goblet cells). Data were normalized to B2MG and ACTB reference genes, and reported as relative expression as compared to GM (set at 1). Results were obtained from 4 different hSIO lines (n = 10–11, Mean ± SD, ***P < 0.001, ****P < 0.0001, Mann Whitney U-test). OLFM4 Olfactomedin, LYZ Lysozyme, I-FABP Intestinal fatty acid-binding protein, MUC2 Mucin 2. C Representative stainings for intestinal protein markers in sections of GM- and DM-cultured hSIO. Proliferation marker Ki67, lysozyme-containing Paneth cells, Alcian Blue-stained Goblet cells, and Alkaline phosphatase staining of the brushborder. Scale bar = 50 µm. D Western blot analysis of lysozyme and IFABP. Band intensity was quantified and normalized using β-actin as loading control. Relative expression is shown as mean ± SD (n = 5, obtained from 3 different hSIO lines; **P < 0.01, Mann Whitney U-test). Full blots can be found in Fig. S1. E Heatmap showing relative change of differentially expressed proteins (P < 0.05). Proteins with on/off expression are excluded from the heatmap and can be found in Table S1. n = 6 from 3 different hSIO lines. F List of known protein markers for intestinal cell-types, with respective abundance ratio (DM/GM) and adjusted p-value. AE absorptive enterocytes, GC Goblet cells, SC stem cells, PC Paneth cells. G Functional enrichment analysis showing GO biological processes significantly enriched (adjusted p-value < 0.05, corresponding to −log(p-value) > 1.3) in DM organoids and GM organoids. Protein lists for enrichment analysis: upregulated in DM p-value < 0.05; upregulated in GM p-value < 0.1. See also Table S1 for the lists of proteins used for enrichment analysis.

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