A Versatile Strategy for Isolating a Highly Enriched Population of Intestinal Stem Cells

• Reporter-free method to purify intestinal stem cells (ISCs)
• Cells share molecular signature with gold standard Lgr5-GFP high cells
• Multidimensional FACS data analysis reveals structure of intestinal crypt
• Lgr5 haploinsufficiency with functional consequences in ISCs from reporter mice

SummaryThe isolation of pure populations of mouse intestinal stem cells (ISCs) is essential to facilitate functional studies of tissue homeostasis, tissue regeneration, and intestinal diseases. However, the purification of ISCs has relied predominantly on the use of transgenic reporter alleles in mice. Here, we introduce a combinational cell surface marker-mediated strategy that allows the isolation of an ISC population transcriptionally and functionally equivalent to the gold standard Lgr5-GFP ISCs. Used on reporter-free mice, this strategy allows the isolation of functional, transcriptionally distinct ISCs uncompromised by Lgr5 haploinsufficiency.

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