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| Item Type: | Article |
|---|---|
| Title: | Isolation of human colon stem cells using surface expression of PTK7 |
| Creators Name: | Jung, P., Sommer, C., Barriga, F.M., Buczacki, S.J., Hernando-Momblona, X., Sevillano, M., Duran-Frigola, M., Aloy, P., Selbach, M., 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 |
| 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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