Fig. 5

Enteroid culture with pre-term metabolites, but not term metabolites, increases proliferation by intestinal stem cell expansion. a MTT assay quantifying enteroid proliferation in presence of term and pre-term metabolites (n = 9 wells per data point pooled from 3 biological replicates). b Representative images showing enteroids cultured with term and pre-term metabolites. c Enteroid area after 3 and 5 days culture with term and pre-term metabolites (n = 20 per group pooled from 2 biological replicates). d Number of crypt domains per enteroid after 3 and 5 days culture with term and pre-term metabolites (n = 20 per group pooled from 2 biological replicates). e Ki67 immunofluorescence in enteroids cultured with term and pre-term stool supernatant. f Quantification of relative Ki67/DAPI signal area in stool supernatant treated enteroids (n = 3 images per group). g Quantitative RT-PCR analysis for markers of proliferation and differentiation in term and pre-term treated enteroids and (h) markers of ER stress and inflammation in term and pre-term treated enteroids (n = 6 per sample pooled from 2 biological replicates). P values calculated using unpaired t-test or Mann-Whitney U tests for all experiments. For all tests, * = P < 0.05, ** = P < 0.01, *** = P < 0.001, control vs. pre-term. # = P < 0.05, ## = P < 0.01, term vs. pre-term