title: Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele.
abstract: 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.
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Onur
authors.initials: O
authors.lastname: Basak
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Maaike
authors.initials: M
authors.lastname: van de Born
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Korving
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Joep
authors.initials: J
authors.lastname: Beumer
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Stefan
authors.initials: S
authors.lastname: van der Elst
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Johan H
authors.initials: JH
authors.lastname: van Es
authors.affiliation: Hubrecht Institute - KNAW and University Medical Centre Utrecht, Utrecht, The Netherlands h.clevers@hubrecht.eu.
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D020869:Gene Expression Profiling; D055879:Gene Knock-In Techniques; D017930:Genes, Reporter; D007413:Intestinal Mucosa / Q000166:cytology / Q000502:physiology; D019394:Ki-67 Antigen / Q000096:biosynthesis / Q000235:genetics; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D046228:Microarray Analysis; D043562:Receptors, G-Protein-Coupled / Q000032:analysis*; D013234:Stem Cells / Q000737:chemistry / Q000502:physiology*; D060449:Wnt Signaling Pathway
chemical_list: D019394:Ki-67 Antigen; C493597:Lgr5 protein, mouse; D008164:Luminescent Proteins; D043562:Receptors, G-Protein-Coupled
keywords: Ki67; Lgr5; intestinal stem cells; quiescent stem cells; secretory lineage
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