Fig. 3: Effects of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) on enterocyte maturation in human preterm intestinal epithelial cells.
From: Functional maturation of preterm intestinal epithelium through CFTR activation
a Schematic illustration of the experimental procedure showing treatment of preterm intestinal epithelial cells with mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs, EV39 and EV42, 2 × 109 EVs/mL for seven days), followed by bulk RNA-seq and qRT-PCR analysis. b Relative proportions of epithelial cell types estimated by deconvolution of bulk RNA-seq data from preterm intestinal epithelial cells treated with MSC-derived EVs (EV39 and EV42, 2 × 109 EVs/mL for seven days), based on previously analyzed scRNA-seq profiles of small intestinal epithelial cells. NT, non-treated group, BEST4, BEST4+ enterocyte; ISC, intestinal stem cell; EEC, enteroendocrine cell; TA, transit-amplifying. c Dot plot showing Z-score transformed relative expression of adult enterocyte marker genes (TMSF20, THSD4, SLC46A1, LCT, ABTB2) in the human preterm intestinal epithelial cells treated with MSC-derived EVs (EV39 and EV42, 2 × 109 EVs/mL) for seven days. NT, non-treated group. d Box plot showing the abundance of age-dependent enterocyte gene sets (first trimester < second trimester < pediatric/adult, n = 224, Supplementary Table 2). NT, non-treated group. Box plot presents minimum, 25% quartile, median, 75% quartile, maximum. e Dot plot showing gene ontology terms obtained from significantly upregulated genes in the preterm intestinal epithelial cells treated with MSC-derived EVs (EV39, 2 × 109 EVs/mL) for seven days compared to non-treated group. Relative mRNA expression of functional enterocyte markers: digestive enzyme genes (f), fatty acid transporter genes (g), amino acid transporter genes (h), inorganic solute transporter genes (i), and cell junction genes (j) in the preterm intestinal epithelial cells treated with MSC-derived EVs (EV39 and EV42, 2 × 109 EVs/mL) for seven days. mRNA level of each gene is normalized to HPRT1 and RPLP0 level. NT, non-treated group (n = 3, 3 wells per group, NT vs. other groups). Quantitative data are expressed as the mean ± standard error of the mean (S.E.M). *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined by one-way ANOVA with Dunnett’s post hoc test (f–j). See also Supplementary Fig. 3.
