An Lgr5-independent developmental lineage is involved in mouse intestinal regeneration II

Collagenase/dispase (CD fraction) treatment of intestinal tissue from adult mice generates cells growing in matrigel as immortal cystic spheroids in addition to differentiated organoids. Contrary to classical EDTA-derived organoids, these spheroids display poor intestinal differentiation and are independent of Rspondin/Noggin/EGF for growth. Their transcriptome resembles strikingly that of fetal intestinal spheroids, with downregulation of crypt base columnar cell (CBC) markers (Lgr5, Ascl2, Smoc2, Olfm4). In addition, they display upregulation of inflammatory and mesenchymal genetic programs, together with robust expression of YAP target genes. Lineage tracing, cell-sorting and single cell RNA sequencing experiments demonstrate that adult spheroid-generating cells belong to a hitherto undescribed developmental lineage, independent of Lgr5+ve CBCs, and are involved in regeneration of the epithelium following CBC ablation Overall design: Examination of the transcriptome of adult intestinal spheroids cultivated in ENR medium upon cells seeding directly in Matrigel.These two sample were compared with small intestinal organoids and fetal small intestinal spheroids presented in an already existed project from our own lab available at GEO under accession number GSE65395. The isolation and the culture conditions remained the same in both studies.