Figure 3.

L. reuteri promotes the secretion of known enteroendocrine-derived intestinal hormones. A) 1) In order to measure the release of intestinal hormones from human intestinal organoids (HIO), L. reuteri conditioned media is generated from mid-log phase cultures of L. reuteri. These cultures are pH neutralized and rendered cell-free. 2) L. reuteri conditioned media is then placed onto NGN3-HIOs plated on transwells that are differentiated but not induced for NGN3 or induced for NGN3. 3) Following an incubation on the HIOs, the supernatant is collected and secreted hormones are measured by ELISA or Luminex assay. Created with BioRender.com. Secreted amylin (B), C-peptide (C), ghrelin (D), GIP (E), PP (F), and PYY (G) measured from uninduced and induced NGN3-HIOs in response to L. reuteri 6475 or 17983 conditioned media. Hormones in B-G were measured on the apical side only of the transwell. In B-G, batches A and B from the RNASeq experiment were pooled so each point on the plot is the result from two organoid batches pooled together. H) Serotonin released from the apical or basolateral side (as indicated) from uninduced and induced NGN3-HIOs in response to L. reuteri 6475 or 17983 conditioned media. In H, shape denotes independent batches of organoids. Only p-values <0.1 are shown with p<0.05 being considered significant. Significance was determined with a Dunnett’s Test.