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使用人类小肠细胞准确定量细胞迁移的优化方法-外文文献论文翻译

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发布于 2017-1-28 22:46:44 | 显示全部楼层 |阅读模式
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外文标题: An optimized method for accurate quantification of cell migration using human small intestine cells
外文摘要: Quantifying the ability of a compound to modulate cell migration rate is a crucial part of many studies including those on chemotaxis, wound healing and cancer metastasis. Existing migration assays all have their strengths and weaknesses. The “scratch” assay is the most widely used because it seems appea- lingly simple and inexpensive. However, the scratch assay has some important limitations, as the tool introducing the “wound” might injure/stress the boundary cells and/or harm underlying matrix coatings, which in both cases will affect cell migration. This described method is a Cell Exclusion Zone Assay, in which cell-free areas are created by growing cells around removable silicone stoppers. Upon appropriate staining with fluorescent dyes and microscopically visualizing the monolayers, the migration rate is then quantified by counting the cells (nuclei) intruding the void area left by the silicone insert. In the current study human small intestine epithelial cells were seeded on a physiological substrate matrix to produce collectively migrating monolayers. Different substrates were tested to determine the optimal surface for enterocyte adherence and migration and morphological changes monitored. Recombinant human epi- dermal growth factor and osteopontin purified from urine were tested to see if the established migration assay produces accurate and reliable migration data with human small intestine cells. The obtained data accurately confirmed that the two bioactive proteins modulate cellular migration in a dose-dependent manner. The presented assay can likely be converted for use with other adherent cell lines or substrate matrices and allows for high throughput, while cost is kept low and versatility high. Co-staining can be applied in order to assay for cell death, different cell types, cell stress and others allowing intricate analysis of migration rate of mixed populations and correction for cell viability.
英关键词: Migration assay Wound healing Collective migration Epithelium Small intestine cells Bioactive
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译文标题: 使用人类小肠细胞准确定量细胞迁移的优化方法
译文概要: -
译文摘要: 量化化合物调节细胞迁移速率的能力是许多研究的重要组成部分,包括趋化性,伤口愈合和癌症转移等主题的研究。现有迁移测定都有其优势和不足。刮伤试验是最普遍的一种,因为操作简单,成本低。但是刮伤试验也有很严重的局限,形成伤口的工具损害或挤压了边缘细胞,又或者破坏基础基质涂层,不论何种情况都会影响细胞迁移。本文方法称为细胞隔离区测验,使细胞生长在可移动硅胶塞周围以形成无细胞区域。适当涂上荧光剂,在显微镜下观察单层,通过计算侵入硅胶插件留下的空隙区域的细胞,量化细胞迁移率。目前的研究将人类小肠上皮细胞接种在生理基质上以获得共同迁移单层。测试不同的底物,确定最适宜肠细胞粘附,迁移和形态学变化的表面。测试从尿液中提纯的重组人表皮生长因子和骨桥蛋白,以确定所建立的使用人类小肠细胞的迁移试验是否形成精确和可靠的迁移数据。获取的数据准确表明两种生物活性蛋白调节细胞迁移且呈现剂量依赖性。加入其他贴壁细胞系或底物基质,现有的试验可转化使用方式,提高吞吐量,而保持成本不变,维持功能多样。多种标记共同使用可以应用于测定细胞死亡,不同的细胞类型,细胞应激等,实现对混合种群迁移率和细胞活力矫正的复杂分析。
译文格式: WORD
中关键词: 迁移试验 伤口愈合 共同迁移 上皮 小肠细胞 生物活性
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