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Isolation of human colon stem cells using surface expression of PTK7

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Item Type:Article
Title:Isolation of human colon stem cells using surface expression of PTK7
Creators Name:Jung, P. and Sommer, C. and Barriga, F.M. and Buczacki, S.J. and Hernando-Momblona, X. and Sevillano, M. and Duran-Frigola, M. and Aloy, P. and Selbach, M. and Winton, D.J. and Batlle, E.
Abstract: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).
Keywords:Cell Adhesion Molecules, Cell Proliferation, Cell Separation, Colon, Cultured Cells, Mass Spectrometry, Organ Culture Techniques, Receptor Protein-Tyrosine Kinases, Stem Cells
Source:Stem Cell Reports
ISSN:2213-6711
Publisher:Cell Press / Elsevier (U.S.A.)
Volume:5
Number:6
Page Range:979-987
Date:8 December 2015
Official Publication:https://doi.org/10.1016/j.stemcr.2015.10.003
PubMed:View item in PubMed

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