FIGURE 3.

Organoids derived from adult intestinal stem cells show intrinsic programming to differentiate into different epithelial cell lineages and express small intestine-specific genes. (A) Cell type-specific transcripts for Crypt Base Columnar (CBC) and Stem cells, Label-retaining (LRC) +4 cells, Proliferation (Prol), Niche factors (NF), Paneth cells (PC), Goblet cells (GC), Enteroendocrine cells (EEC), and Absorptive cells or enterocytes (Abs/EC). (B) Overlaying the NCBI gene tracks of MUC2 NC_010444.4 on chromosome 2 at location 689363–719542bp (green area), shows identical overlap with the mapped reads and coverage (Cov) from organoid and tissue samples, but not in IPEC-J2 (ENS; Ensembl reference genome). To confirm MUC2 protein translation and subsequent mucus formation, Carnoy fixed tissue, organoid, and IPEC-J2 samples were stained with PAS/Alcian blue (left) and porcine anti-MUC2 (right; black and white size bars indicate 100 μm). (C) Most small intestine-specific genes identified in the human protein atlas are also expressed in porcine jejunum tissue and their derived organoids (>74%), whereas fewer are expressed in ileum organoids (52%). IPEC-J2 only expressed 32% of the small intestine-specific genes. (D) Organoid transcriptomes from jejunum and ileum were compared to identify differences in expressed genes, showing large overlap of genes (Venn), but also some differences. The different genes were analyzed using TOPPfun to identify putative differences in gene ontology and pathways (TMT, Transmembrane transport; PSP, Peptidyl-serine phosphorylation).