Abstract
Collagenase/dispase treatment of intestinal tissue from adult mice generates cells growing in matrigel as stably replatable 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.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
This version of the manuscript has been revised taking into account the points raised by the three referees to whom the previous version had been submitted by Review Commons. It contains additional data in the form of an additional supplementary Table (Table S2, former Table S2 becoming Table S3). Please note that additional results answering selected points raised by referees but not included in the manuscript are presented in the supplementary file "revision-plan-RC-2024-02491.pdf"
