An ex vivo assay to assess the potential of skin keratinocytes for wound epithelialization
An ex vivo assay to assess the potential of skin keratinocytes for wound epithelialization
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DOI：
10.1046/j.1523-1747.2002.01736.x
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发表时间：
2002-05-01
影响因子：
6.5
通讯作者：
Coulombe, PA
中科院分区：
文献类型：
--
作者：
Mazzalupo, S;Wawersik, MJ;Coulombe, PA
皮肤损伤后的伤口闭合是一个复杂的过程，涉及真皮收缩和角质形成细胞迁移。创伤愈合的鼠类模型是潜在有用的，因为能够通过基因操作来确定蛋白质功能。由于皮肤收缩的主导作用，在准备伤口部位进行形态学研究的技术困难，以及出生后改变转基因皮肤特性的表型，在评估上皮细胞对小鼠皮肤伤口闭合的贡献方面存在困难。我们描述了一个简单的体外试验，利用外植体培养，使定量评估的潜力，小鼠角质形成细胞的伤口上皮化。在该测定中，皮肤角质形成细胞的行为和性质很好地模拟了原位皮肤伤口边缘处发生的行为和性质，包括对结缔组织成分、增殖和迁移的依赖性。可以实时研究或在特定时间点检查从皮肤外植体出现的上皮细胞生长，以获得上皮形成过程中的感兴趣的标志物。该测定是定量的，并且可以成功地检测上皮形成潜力的增加或减少，并且可以用于转基因小鼠模型的表征。
Wound closure following injury to the skin is a complex process involving both dermal contraction and keratinocyte migration. Murine models of wound healing are potentially useful because of the ability to determine protein function through gene manipulation. Owing to the dominant role of dermal contraction, the technical difficulties in preparing the wound site for morphologic studies, and the postnatal phenotypes altering the properties of transgenic skin, there are difficulties in assessing the epithelial contribution to wound closure in mouse skin. We describe a simple ex vivo assay utilizing explant culture that enables a quantitative assessment of the potential of mouse keratinocytes for wound epithelialization. In this assay, the behavior and properties of skin keratinocytes mimic well those that occur at the edge of skin wounds in situ , including a dependence upon connective tissue element(s), proliferation, and migration. The epithelial cell outgrowths emerging from skin explants can be studied in real-time or examined at specific time-points for markers of interest in the epithelialization process. The assay is quantitative and can successfully detect increases or decreases in epithelialization potential, and can be useful in the characterization of transgenic mouse models.