Characterization and Localization of Side Population Cells in Mouse Skin
Characterization and Localization of Side Population Cells in Mouse Skin
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DOI：
10.1634/stemcells.2004-0226
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发表时间：
2005-06
期刊：
影响因子：
5.2
通讯作者：
S. Yano;Yuriko Ito;M. Fujimoto;T. Hamazaki;K. Tamaki;H. Okochi
中科院分区：
文献类型：
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作者：
S. Yano;Yuriko Ito;M. Fujimoto;T. Hamazaki;K. Tamaki;H. Okochi
近年来，侧群(SP)细胞的检测作为干细胞分离的一种方法引起了人们的关注，SP细胞具有很强的外排Hoechst 33342荧光染料的能力。我们从小鼠皮肤中鉴定SP细胞的方法与从骨髓中鉴定的方法相同。在钙通道阻滞剂维拉帕米治疗后，这一群体几乎完全消失。SP细胞主要分布在表皮，真皮中有少量分布。随着年龄的增长，SP细胞的比例下降。表面标记分析显示，分离的SP细胞高表达α6整合素、β1整合素、Sca 1、角蛋白14和角蛋白19，而低水平表达E钙粘附素、CD34和CD71。在分离的SP细胞中，参与染料外流的乳腺癌耐药蛋白1(BCRP1)在蛋白质和mRNA水平均有高水平的表达。免疫组织化学分析表明，BCRP1在小鼠皮肤基底层和毛发隆起区均有表达。原位杂交结果显示，BCRP1基因在新生皮肤的基底层和毛囊中均有表达。这些结果表明，BCRP1阳性细胞的定位与角质形成干细胞的定位是一致的。根据BCRP1与SP细胞表型的密切关系，我们认为角质形成细胞干细胞与SP或BCRP1阳性细胞关系密切。
Recently, the detection of side population (SP) cells, which have the ability to strongly efflux Hoechst 33342 fluorescence dye, has attracted attention as a method of stem cell isolation. We identified SP cells from mouse skin using the same method as from bone marrow. This population almost completely disappeared after treatment with the calcium channel blocker verapamil. SP cells were mainly localized in the epidermis, with a few in the dermis. The ratio of SP cells decreased as the mouse became older. Surface marker analysis revealed that the sorted SP cells expressed α6‐integrin, β1‐integrin, Sca‐1, keratin 14, and keratin 19, which are proliferating and progenitor cell markers, at levels higher than in non‐SP cells, while they expressed E‐cadherin, CD34, and CD71 at lower levels. The expression of breast cancer resistance protein 1 (BCRP1), which participates in dye efflux, was expressed at high levels at both the protein and mRNA level in sorted SP cells. Immunohistochemical analysis showed that BCRP1 was expressed in the basal layers and hair bulge regions of mouse skin. BCRP1 mRNA was found in basal layers and hair follicles of newborn skin by in situ hybridization. These results indicate that the localization of BCRP1‐positive cells is compatible with that of keratinocyte stem cells. Based on the close relationship between BCRP1 and the SP cell phenotype, we conclude that keratinocyte stem cells are closely related to the SP‐ or BCRP1‐positive cells.