title: Isolation of Human Colon Stem Cells Using Surface Expression of PTK7.
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).
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
authors.forename: Peter
authors.initials: P
authors.lastname: Jung
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13092 Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Sommer
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
authors.forename: Francisco M
authors.initials: FM
authors.lastname: Barriga
authors.affiliation: Cancer Research UK, Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Buczacki
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Hernando-Momblona
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Sevillano
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain.
authors.forename: Miquel
authors.initials: M
authors.lastname: Duran-Frigola
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain.
authors.forename: Patrick
authors.initials: P
authors.lastname: Aloy
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13092 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
authors.affiliation: Cancer Research UK, Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, UK.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Winton
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, 08028 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain. Electronic address: eduard.batlle@irbbarcelona.org.
authors.forename: Eduard
authors.initials: E
authors.lastname: Batlle
mesh_terms: D015815:Cell Adhesion Molecules / Q000032:analysis*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D013058:Mass Spectrometry; D009924:Organ Culture Techniques; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis*; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology*
chemical_list: D015815:Cell Adhesion Molecules; C096905:PTK7 protein, human; D020794:Receptor Protein-Tyrosine Kinases