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利用磁珠细胞分选法(MACS)分离成年小鼠干细胞角质形成细胞。

Isolation of adult mouse stem keratinocytes using magnetic cell sorting (MACS).

作者信息

Lorz Corina, Segrelles Carmen, Garín Marina, Paramio Jesús M

机构信息

Molecular Oncology Unit, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Madrid, Spain.

出版信息

Methods Mol Biol. 2010;585:1-11. doi: 10.1007/978-1-60761-380-0_1.

Abstract

The functional and molecular characterization of mouse epidermal stem cells (SC) requires in many circumstances arduous technologies and elaboration of complicated transgenic mouse models. To avoid these difficulties, we have developed a simplified protocol to obtain primary keratinocytes from adult mouse skin followed by magnetic-coupled isolation of the CD34 positive epidermal SC population. This approach provides a reproducible and useful method to obtain large amounts of viable cells in a quick and affordable manner, which allows the analysis of pathologies or mutant mice with reduced epidermal SC, and for the molecular and genomic analysis of this population.

摘要

在许多情况下,对小鼠表皮干细胞(SC)进行功能和分子特征分析需要采用艰巨的技术,并构建复杂的转基因小鼠模型。为避免这些困难,我们开发了一种简化方案,先从成年小鼠皮肤中获取原代角质形成细胞,然后通过磁耦合分离出CD34阳性表皮干细胞群体。这种方法提供了一种可重复且实用的方式,能够快速且经济高效地获取大量活细胞,这有助于分析表皮干细胞数量减少的病理情况或突变小鼠,并对该细胞群体进行分子和基因组分析。

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