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Isolation of human colon stem cells using surface expression of PTK7

Authors

  • P. Jung
  • C. Sommer
  • F.M. Barriga
  • S.J. Buczacki
  • X. Hernando-Momblona
  • M. Sevillano
  • M. Duran-Frigola
  • P. Aloy
  • M. Selbach
  • D.J. Winton
  • E. Batlle

Journal

  • Stem Cell Reports

Citation

  • Stem Cell Rep 5 (6): 979-987

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).


DOI

doi:10.1016/j.stemcr.2015.10.003