Isolation of Human Colon Stem Cells Using Surface Expression of PTK7.
Isolation of Human Colon Stem Cells Using Surface Expression of PTK7.
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使用PTK7的表面表达分离人类结肠干细胞。
DOI：
10.1016/j.stemcr.2015.10.003
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发表时间：
2015-12-08
影响因子：
5.9
通讯作者：
Batlle E
中科院分区：
文献类型：
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作者：
Jung P;Sommer C;Barriga FM;Buczacki SJ;Hernando-Momblona X;Sevillano M;Duran-Frigola M;Aloy P;Selbach M;Winton DJ;Batlle E
将报告基因盒插入Lgr5基因座中能够表征小鼠肠干细胞（ISCs）。然而，LGR 5蛋白的低细胞表面丰度和缺乏高亲和力抗LGR 5抗体代表了有效分离人结肠干细胞（hCoSC）的障碍。我们着手鉴定允许纯化hCoSC的干细胞标志物。在无偏方法中，通过定量质谱法分析源自体外人结肠类器官的膜富集蛋白质级分。蛋白酪氨酸假激酶PTK7在人类结肠类器官中指定了一个细胞群，其特征在于最高的自我更新和重新接种能力。识别PTK7胞外结构域的抗体使我们能够直接从患者来源的粘膜样品中分离和扩增hCoSC。人PTK7+细胞显示典型Lgr5+ ISC的特征，并且包括经历向肠内分泌谱系分化的一部分细胞，其类似于隐窝标记保留细胞（LRC）。离体培养的人结肠干细胞（hCoSC）表达表面标志物PTK7具有最高PTK7表面丰度的hCoSC显示最高的自我更新能力抗PTK7的抗体能够从患者样品中纯化hCoSC PTK7+ hCoSC的一部分显示隐窝标记保留细胞的遗传特征在这篇文章中，Batlle及其同事表明，蛋白质假激酶7（PTK7）代表了人类结肠干细胞（hCoSC）的高度特异性标志物。PTK7表面丰度可用于从体外结肠类器官培养物和患者来源的粘膜样品富集hCoSC。PTK7+ hCoSC组分显示典型Lgr5+肠干细胞和慢循环隐窝标记保留细胞（LRC）的遗传特征。
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). Ex vivo cultured human colon stem cells (hCoSCs) express the surface marker PTK7 hCoSCs with highest PTK7 surface abundance display highest self-renewal capacity Antibodies against PTK7 enable purification of hCoSCs from patient samples A fraction of PTK7+ hCoSCs display genetic features of crypt label retaining cells In this article, Batlle and colleagues show that the protein pseudo-kinase 7 (PTK7) represents a highly specific marker for human colonic stem cells (hCoSCs). PTK7 surface abundance can be used to enrich for hCoSCs from in vitro colonic organoid cultures and patient-derived mucosa samples. PTK7+ hCoSC fractions display genetic features of canonical Lgr5+ intestinal stem cells and slow cycling crypt label retaining cells (LRCs).