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Insertion of reporter cassettes into the Lgr5 locus has enabled the characterization of mouse intestinal stem cells (ISCs). However, low cell surface abundance of LGR5 protein and lack of high-affinity anti-LGR5 antibodies represent a roadblock to efficiently isolate human colonic stem cells (hCoSCs). We set out to identify stem cell markers that would allow for purification of hCoSCs. In an unbiased approach, membrane-enriched protein fractions derived from in vitro human colonic organoids were analyzed by quantitative mass spectrometry. Protein tyrosine pseudokinase PTK7 specified a cell population within human colonic organoids characterized by highest self-renewal and re-seeding capacity. Antibodies recognizing the extracellular domain of PTK7 allowed us to isolate and expand hCoSCs directly from patient-derived mucosa samples. Human PTK7+ cells display features of canonical Lgr5+ ISCs and include a fraction of cells that undergo differentiation toward enteroendocrine lineage that resemble crypt label retaining cells (LRCs).

Original publication

DOI

10.1016/j.stemcr.2015.10.003

Type

Journal article

Journal

Stem Cell Reports

Publication Date

08/12/2015

Volume

5

Pages

979 - 987

Keywords

Cell Adhesion Molecules, Cell Proliferation, Cell Separation, Cells, Cultured, Colon, Humans, Mass Spectrometry, Organ Culture Techniques, Receptor Protein-Tyrosine Kinases, Stem Cells