Enrichment for murine keratinocyte stem cells based on cell surface phenotype
Enrichment for murine keratinocyte stem cells based on cell surface phenotype
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DOI：
10.1073/pnas.97.20.10960
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发表时间：
2000-09-26
影响因子：
11.1
通讯作者：
Kaur, P
中科院分区：
文献类型：
--
作者：
Tani, H;Morris, RJ;Kaur, P
上皮干细胞的识别和物理分离对于我们了解其稳态过程中的生长调节至关重要。伤口愈合和癌变。这些干细胞的特征仍然很差，因为缺乏特定的分子标记，使我们能够将它们与其后代、转运放大（TA）细胞区分开来，后者的增殖潜力更有限。细胞动力学分析允许将小鼠角质形成细胞干细胞 (KSC) 鉴定为保留 [H-3] 胸苷 ([H-3]Tdr) 标记的缓慢循环细胞，称为标记保留细胞 (LRC)，而 TA 细胞在 [H-3]Tdr 单脉冲后可视化为快速循环细胞。称为脉冲标记细胞（PLC），在这里，我们报告了通过结合使用体内细胞动力学分析和荧光激活细胞分选，成功地将 KSC 与 TA 细胞分离。具体来说，我们证明，小鼠背部角质形成细胞的特征是高水平的α（6）整合素和低至不可检测的转铁蛋白受体（CD71）表达，称为α（6）（bri）CD71（dim）细胞，富含上皮干细胞，因为它们代表了小胚细胞样细胞的少量（约8％）和静止亚群。具有高核：细胞质比率，含有大约 70% 的标记保留细胞，后者是干细胞的一个有据可查的特征。反过来。 TA 细胞可以富集在表型不同的亚群中，称为 alpha(6)(bri)CD71(dim)。代表大部分（约 60%）活跃循环的基底角质形成细胞，重要的是含有约 70% 的 [H-3]Tdr 脉冲标记细胞。重要的是，背部皮肤的免疫染色显示毛囊隆起区域存在 CD71(dim)，这是 KSC 的一个有据可查的位置。
The identification and physical isolation of epithelial stem cells is critical to our understanding of their growth regulation during homeostasis. wound healing, and carcinogenesis. These stem cells remain poorly characterized because of the absence of specific molecular markers that permit us to distinguish them from their progeny, the transit amplifying (TA) cells, which have a more restricted proliferative potential. Cell kinetic analyses have permitted the identification of murine keratinocyte stem cells (KSCs) as slowly cycling cells that retain [H-3]thymidine ([H-3]Tdr) label, termed label-retaining cells (LRCs), whereas TA cells are visualized as rapidly cycling cells after a single pulse of [H-3]Tdr. termed pulse-labeled cells (PLCs), Here, we report on the successful separation of KSCs from TA cells through the combined use of in vivo cell kinetic analysis and fluorescence-activated cell sorting. Specifically, we demonstrate that murine dorsal keratinocytes characterized by their high levels of alpha(6) integrin and low to undetectable expression of the transferrin receptor (CD71) termed alpha(6)(bri)CD71(dim) cells, are enriched for epithelial stem cells because they represent a minor (approximate to 8%) and quiescent subpopulation of small blast-like cells. with a high nuclear:cytoplasmic ratio, containing approximate to 70% of label-retaining cells, the latter being a well documented characteristic of stem cells. Conversely. TA cells could be enriched in a phenotypically distinct subpopulation termed alpha(6)(bri)CD71(dim). representing the majority (approximate to 60%) of basal keratinocytes that are actively cycling, and importantly contain approximate to 70% of [H-3]Tdr pulse-labeled cells. Importantly, immunostaining of dorsal skin revealed the presence of CD71(dim) tells in the hair follicle bulge region, a well documented location for KSCs.