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首页> 外文期刊>Cellular and Molecular Bioengineering >Label-Free Automated Cell Tracking: Analysis of the Role of E-cadherin Expression in Collective Electrotaxis
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Label-Free Automated Cell Tracking: Analysis of the Role of E-cadherin Expression in Collective Electrotaxis

机译：无标记自动细胞追踪：E-钙粘蛋白表达在集体趋电性中的作用分析

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摘要

Collective cell migration plays an important role in wound healing, organogenesis, and the progression of metastatic disease. Analysis of collective migration typically involves laborious and time-consuming manual tracking of individual cells within cell clusters over several dozen or hundreds of frames. Herein, we develop a label-free, automated algorithm to identify and track individual epithelial cells within a free-moving cluster. We use this algorithm to analyze the effects of partial E-cadherin knockdown on collective migration of MCF-10A breast epithelial cells directed by an electric field. Our data show that E-cadherin knockdown in free-moving cell clusters diminishes electrotactic potential, with empty vector MCF-10A cells showing 16 higher directedness than cells with E-cadherin knockdown. Decreased electrotaxis is also observed in isolated cells at intermediate electric fields, suggesting an adhesion-independent role of E-cadherin in regulating electrotaxis. In additional support of an adhesion-independent role of E-cadherin, isolated cells with reduced E-cadherin expression reoriented within an applied electric field 60 more quickly than control. These results have implications for the role of E-cadherin expression in electrotaxis and demonstrate proof-of-concept of an automated algorithm that is broadly applicable to the analysis of collective migration in a wide range of physiological and pathophysiological contexts.
机译：集体细胞迁移在伤口愈合、器官发生和转移性疾病的进展中起着重要作用。集体迁移的分析通常涉及在几十或数百帧内对细胞簇中的单个细胞进行费力且耗时的手动跟踪。在这里，我们开发了一种无标记的自动化算法来识别和跟踪自由移动簇中的单个上皮细胞。我们使用该算法分析部分 E-钙粘蛋白敲低对电场引导的 MCF-10A 乳腺上皮细胞集体迁移的影响。我们的数据表明，自由移动的细胞簇中的E-钙粘蛋白敲低会降低电趋向电位，空载体MCF-10A细胞的定向性比E-钙粘蛋白敲低的细胞高16%。在中间电场的分离细胞中也观察到趋电性降低，表明 E-钙粘蛋白在调节趋电性中具有不依赖粘附的作用。为了进一步支持E-钙粘蛋白的粘附非依赖性作用，E-钙粘蛋白表达降低的分离细胞在施加的电场中的重新定向速度比对照组快60%。这些结果对E-钙粘蛋白表达在趋电性中的作用具有重要意义，并证明了一种自动化算法的概念验证，该算法广泛适用于在广泛的生理和病理生理学背景下分析集体迁移。

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