Label-Free Automated Cell Tracking: Analysis of the Role of E-cadherin Expression in Collective Electrotaxis
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- 作者：Mark L. Lalli ; Brooke Wojeski ; Anand R. Asthagiri
- 关键词：Cell–cell interactions ; Electrotaxis ; Guidance cues ; Image analysis
- 刊名：Cellular and Molecular Bioengineering
- 出版年：2017
- 出版时间：February 2017
- 年：2017
- 卷：10
- 期：1
- 页码：89-101
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- 刊物类别：Engineering
- 刊物主题：Biomedical Engineering; Classical Mechanics; Continuum Mechanics and Mechanics of Materials; Biological and Medical Physics, Biophysics; Biomaterials; Cell Biology;
- 出版者：Springer US
- ISSN：1865-5033
- 卷排序：10
文摘
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.