Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele

Cycling Lgr5+ stem cells fuel the rapid turnover of the adult intestinal epithelium. The existence of quiescent Lgr5+ cells has been reported, while an alternative quiescent stem cell population is believed to reside at crypt position +4. Here, we generated a novel Ki67RFP knock-in allele that ident...

Full description

Bibliographic Details
Main Authors:	Basak, Onur, van de Born, Maaike, Korving, Jeroen, Beumer, Joep, van der Elst, Stefan, van Es, Johan H, Clevers, Hans
Format:	Online
Language:	English
Published:	BlackWell Publishing Ltd 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4195772/

id	pubmed-4195772
recordtype	oai_dc
spelling	pubmed-41957722015-09-17 Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele Basak, Onur van de Born, Maaike Korving, Jeroen Beumer, Joep van der Elst, Stefan van Es, Johan H Clevers, Hans Articles Cycling Lgr5+ stem cells fuel the rapid turnover of the adult intestinal epithelium. The existence of quiescent Lgr5+ cells has been reported, while an alternative quiescent stem cell population is believed to reside at crypt position +4. Here, we generated a novel Ki67RFP knock-in allele that identifies dividing cells. Using Lgr5-GFP;Ki67RFP mice, we isolated crypt stem and progenitor cells with distinct Wnt signaling levels and cell cycle features and generated their molecular signature using microarrays. Stem cell potential of these populations was further characterized using the intestinal organoid culture. We found that Lgr5high stem cells are continuously in cell cycle, while a fraction of Lgr5low progenitors that reside predominantly at +4 position exit the cell cycle. Unlike fast dividing CBCs, Lgr5low Ki67− cells have lost their ability to initiate organoid cultures, are enriched in secretory differentiation factors, and resemble the Dll1 secretory precursors and the label-retaining cells of Winton and colleagues. Our findings support the cycling stem cell hypothesis and highlight the cell cycle heterogeneity of early progenitors during lineage commitment. BlackWell Publishing Ltd 2014-09-17 2014-08-04 /pmc/articles/PMC4195772/ /pubmed/25092767 http://dx.doi.org/10.15252/embj.201488017 Text en © 2014 The Authors. Published under the terms of the CC BY NC ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Basak, Onur van de Born, Maaike Korving, Jeroen Beumer, Joep van der Elst, Stefan van Es, Johan H Clevers, Hans
spellingShingle	Basak, Onur van de Born, Maaike Korving, Jeroen Beumer, Joep van der Elst, Stefan van Es, Johan H Clevers, Hans Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
author_facet	Basak, Onur van de Born, Maaike Korving, Jeroen Beumer, Joep van der Elst, Stefan van Es, Johan H Clevers, Hans
author_sort	Basak, Onur
title	Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
title_short	Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
title_full	Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
title_fullStr	Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
title_full_unstemmed	Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele
title_sort	mapping early fate determination in lgr5+ crypt stem cells using a novel ki67-rfp allele
description	Cycling Lgr5+ stem cells fuel the rapid turnover of the adult intestinal epithelium. The existence of quiescent Lgr5+ cells has been reported, while an alternative quiescent stem cell population is believed to reside at crypt position +4. Here, we generated a novel Ki67RFP knock-in allele that identifies dividing cells. Using Lgr5-GFP;Ki67RFP mice, we isolated crypt stem and progenitor cells with distinct Wnt signaling levels and cell cycle features and generated their molecular signature using microarrays. Stem cell potential of these populations was further characterized using the intestinal organoid culture. We found that Lgr5high stem cells are continuously in cell cycle, while a fraction of Lgr5low progenitors that reside predominantly at +4 position exit the cell cycle. Unlike fast dividing CBCs, Lgr5low Ki67− cells have lost their ability to initiate organoid cultures, are enriched in secretory differentiation factors, and resemble the Dll1 secretory precursors and the label-retaining cells of Winton and colleagues. Our findings support the cycling stem cell hypothesis and highlight the cell cycle heterogeneity of early progenitors during lineage commitment.
publisher	BlackWell Publishing Ltd
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4195772/
_version_	1613144291868147712

Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele

Similar Items