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Intestinal label-retaining cells are secretory precursors expressing Lgr5

Abstract

The rapid cell turnover of the intestinal epithelium is achieved from small numbers of stem cells located in the base of glandular crypts. These stem cells have been variously described as rapidly cycling or quiescent. A functional arrangement of stem cells that reconciles both of these behaviours has so far been difficult to obtain. Alternative explanations for quiescent cells have been that they act as a parallel or reserve population that replace rapidly cycling stem cells periodically or after injury; their exact nature remains unknown. Here we show mouse intestinal quiescent cells to be precursors that are committed to mature into differentiated secretory cells of the Paneth and enteroendocrine lineage. However, crucially we find that after intestinal injury they are capable of extensive proliferation and can give rise to clones comprising the main epithelial cell types. Thus, quiescent cells can be recalled to the stem-cell state. These findings establish quiescent cells as an effective clonogenic reserve and provide a motivation for investigating their role in pathologies such as colorectal cancers and intestinal inflammation.

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Figure 1: Identification and isolation of YFP–LRCs.
Figure 2: YFP–LRCs are a discrete Lgr5-expressing subpopulation.
Figure 3: Acquisition of Paneth and enteroendocrine cell characteristics from YFP–LRCs.
Figure 4: LRCs only demonstrate clonogenicity after injury in vivo.

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Gene Expression Omnibus

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Data were deposited to the GEO database under accession number GSE43772.

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Acknowledgements

This research was supported by Cancer Research UK (S.J.A.B., H.I.Z., A.M.N., R.R., R.K. and D.J.W.). L.V. was supported by a KWF fellowship. We thank D. Tan for providing the single-cell RNA amplification protocol. We also thank M. de la Roche, R. von Furstenberg and S. Henning for advice with the in vitro culture work. We acknowledge the following core facilities at CRUK/CRI: The Transgenic Laboratory, Biological Resource Unit, Flow Cytometry, Histopathology, Microscopy, Genomics and Bioinformatics and the CRUK Paterson Institute Microarray Facility. We thank R. J. Davies, A. Klein and A. Ibrahim for manuscript discussions.

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Authors

Contributions

S.J.A.B. designed and performed experiments and wrote the paper. H.I.Z. designed and developed the H2B–YFP model and performed experiments. R.R. performed the bioinformatic analysis. A.M.N. and L.V. performed experiments. R.K. designed experiments and performed bioinformatic analysis. D.J.W. designed experiments, developed the diCreAB model and wrote the paper.

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Correspondence to Douglas J. Winton.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-6. (PDF 2399 kb)

H2B-YFP Paneth cells are non-responsive to βNF induction

Confocal reconstruction video of an isolated crypt immediately following βNF treatment, showing all cells other than Paneth cells express YFP. Yellow=YFP. Red=UEA. (MOV 4370 kb)

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Buczacki, S., Zecchini, H., Nicholson, A. et al. Intestinal label-retaining cells are secretory precursors expressing Lgr5. Nature 495, 65–69 (2013). https://doi.org/10.1038/nature11965

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