Premium
Mapping early fate determination in L gr5 + crypt stem cells using a novel K i67‐ RFP allele
Author(s) -
Basak Onur,
Born Maaike,
Korving Jeroen,
Beumer Joep,
Elst Stefan,
Es Johan H,
Clevers Hans
Publication year - 2014
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201488017
Subject(s) - lgr5 , stem cell , biology , progenitor cell , microbiology and biotechnology , crypt , wnt signaling pathway , stem cell marker , organoid , cell cycle , population , adult stem cell , cellular differentiation , immunology , cell , cancer stem cell , genetics , signal transduction , endocrinology , gene , demography , sociology
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 Ki67 RFP knock‐in allele that identifies dividing cells. Using Lgr5‐ GFP ;Ki67 RFP 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 Lgr5 high stem cells are continuously in cell cycle, while a fraction of Lgr5 low progenitors that reside predominantly at +4 position exit the cell cycle. Unlike fast dividing CBC s, Lgr5 low 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.