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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).

journal_name

Stem Cell Reports

journal_title

Stem cell reports

authors

Jung P,Sommer C,Barriga FM,Buczacki SJ,Hernando-Momblona X,Sevillano M,Duran-Frigola M,Aloy P,Selbach M,Winton DJ,Batlle E

doi

10.1016/j.stemcr.2015.10.003

subject

Has Abstract

pub_date

2015-12-08 00:00:00

pages

979-987

issue

6

issn

2213-6711

pii

S2213-6711(15)00300-8

journal_volume

5

pub_type

杂志文章

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