Fig. 4

Intraperitoneal administration of thMSC-EVs increased goblet cells and ISC marker gene expression in NEC-induced mouse pups.(A) Bar graph representation of the expression levels of intestinal marker genes LGR5, OLFM4, ATOH1, and FABP6 analyzed via RT-qPCR. n = 16, 11, and 16 in the CON, NEC, and NEC + thEV groups, respectively. The Mann–Whitney test was used for the analysis. (B) Bar graph representation of the count of TUNEL (+) cells. n = 11, 21, and 25 in the CON, NEC, and NEC + thEV groups, respectively. One-way ANOVA and the post-hoc Bonferroni test were used in the analysis. (C) Bar graph representation of pro-inflammatory cytokine IL-1α levels measured using ELISA. n = 18, 23, and 19 in the CON, NEC, and NEC + EV groups, respectively. (D) Representative microscopic image of immunofluorescent-stained OLFM4 (+) cells and TUNEL (+) cells. (E) Representative microscopic image of merged OLFM4 and TUNEL (+) cells. One-way ANOVA and the post-hoc Bonferroni test were used in the analysis. White arrows point to TUNEL (+) cells. Data are presented as mean ± SEM. *, p < 0.01 vs. CON; #, p < 0.01 vs. NEC. CON, normal control; NEC, necrotizing enterocolitis control group; NEC + thEV, thMSC-EVs treated NEC group