Isolation of Human Colon Stem Cells Using Surface Expression of PTK7.
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Authors
Jung, Peter
Sommer, Christian
Barriga, Francisco M
Hernando-Momblona, Xavier
Sevillano, Marta
Duran-Frigola, Miquel
Aloy, Patrick
Selbach, Matthias
Batlle, Eduard
Publication Date
2015-12-08Journal Title
Stem Cell Reports
ISSN
2213-6711
Publisher
Elsevier BV
Volume
5
Issue
6
Pages
979-987
Language
eng
Type
Article
Physical Medium
Print-Electronic
Metadata
Show full item recordCitation
Jung, P., Sommer, C., Barriga, F. M., Buczacki, S., Hernando-Momblona, X., Sevillano, M., Duran-Frigola, M., et al. (2015). Isolation of Human Colon Stem Cells Using Surface Expression of PTK7.. Stem Cell Reports, 5 (6), 979-987. https://doi.org/10.1016/j.stemcr.2015.10.003
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
Colon, Cells, Cultured, Stem Cells, Humans, Receptor Protein-Tyrosine Kinases, Cell Adhesion Molecules, Organ Culture Techniques, Cell Separation, Cell Proliferation, Mass Spectrometry
Sponsorship
Cancer Research UK (C14303/A17197)
Identifiers
External DOI: https://doi.org/10.1016/j.stemcr.2015.10.003
This record's URL: https://www.repository.cam.ac.uk/handle/1810/279437
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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