Magnetic separation and characterization of keratinocyte stem cells from human gingiva
Magnetic separation and characterization of keratinocyte stem cells from human gingiva
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DOI：
10.1111/j.1600-0765.2010.01284.x
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发表时间：
2010-12-01
影响因子：
3.5
通讯作者：
Hirata, T.
中科院分区：
文献类型：
--
作者：
Calenic, B.;Ishkitiev, N.;Hirata, T.
背景和目的：尽管角质形成干细胞在组织稳态、伤口愈合和肿瘤形成中起着关键作用，但它们仍然难以识别和表征。本研究的目的是分离和表征口腔角质形成细胞干细胞population.Material和方法：口腔人角质形成细胞角化口腔粘膜获得磁性分离使用α（6）β（4）整合素和增殖相关的标志物，CD 71。分析分离的细胞级分的细胞大小、细胞周期阶段（使用流式细胞术）和集落形成能力。结果：干细胞α（6）β（阳性）（4）CD 71（阴性）部分与α（6）β（阳性）（4）CD 71（阳性）和α（6）β 4（阴性）部分相比具有最小的细胞大小[分别为780.7 +/- 141.5（像素），1422.9 +/- 264.6（像素）和3844.4 +/- 220.1（像素），p < 0.01;方差分析（ANOVA）]。此外，α（4）β（阳性）（4）CD 71（阴性）亚群在三种细胞组分中始终具有最高的集落形成能力（分别与alpha（6）beta（pos）（4）CD 71（pos）和alpha（6）beta（neg）（4）亚群相比，126.2 +/- 21.7 vs. 32.8 +/- 4.5 vs. 12.4 +/- 2.1，p < 0.01;方差分析）。此外，与α（6）β（阳性）（4）CD 71（阳性）细胞组分相比，α（6）β（阳性）（4）CD 71（阴性）组分含有更多的静止细胞和更少的活跃循环细胞。候选干细胞的细胞角蛋白19和p63角质形成细胞干细胞标记阳性，而分化标记，如细胞角蛋白10或involucrin是absent.Conclusion：人牙龈α（6）β（pos）（4）CD 71（neg）细胞馏分，分离的磁性系统，表现出牙龈角质形成细胞干细胞的几个特点。也有人建议，磁系统可能是一个重要的工具，在获取口腔角质形成干细胞的研究。
Background and Objective:Although keratinocyte stem cells play a key role in tissue homeostasis, wound healing and neoplasia, they remain difficult to identify and characterize. The purpose of this study was to isolate and characterize an oral keratinocyte stem-cell population.Material and Methods:Oral human keratinocytes obtained from keratinized oral mucosa were magnetically separated using alpha(6)beta(4) integrin and a proliferation-related marker, CD71. The isolated cell fractions were analyzed for cell size, cell cycle stage (using flow cytometry) and colony-forming ability. The expression of stem cell markers p63 and cytokeratin 19 and of differentiation markers cytokeratin 10 and involucrin was checked using immunocytochemical analysis.Results:The stem cell alpha(6)beta(pos)(4) CD71(neg) fraction had the smallest cell size compared with alpha(6)beta(pos)(4) CD71(pos) and alpha(6)beta 4(neg) fractions [ 780.7 +/- 141.5 ( pixels), 1422.9 +/- 264.6 ( pixels) and 3844.4 +/- 220.1 (pixels) respectively, p < 0.01; analysis of variance ( ANOVA)]. Also, the alpha(4)beta(pos)(4)CD71(neg) subpopulation consistently had the highest colony- forming ability among the three cell fractions ( 126.2 +/- 21.7 vs. 32.8 +/- 4.5 vs. 12.4 +/- 2.1 compared with alpha(6)beta(pos)(4) CD71(pos) and alpha(6)beta(neg)(4) subpopulations, respectively, p < 0.01; ANOVA). Moreover, the alpha(6)beta(pos)(4) CD71(neg) fraction contained more quiescent cells and fewer actively cycling cells than the alpha(6)beta(pos)(4) CD71(pos) cell fraction. The candidate stem cells were positive for cytokeratin 19 and p63 keratinocyte stem cell markers, while differentiation markers such as cytokeratin 10 or involucrin were absent.Conclusion:The human gingival alpha(6)beta(pos)(4) CD71(neg) cell fraction, separated by a magnetic system, demonstrated several characteristics of gingival keratinocyte stem cells. It is also suggested that a magnetic system may be an important tool in acquiring oral keratinocyte stem cells for research.