Protocol for electrotaxis of large epithelial cell sheets

• Published in: STAR protocols Vol. 4; no. 2; p. 102288
• Main Authors: Zhang, Yan, Lee, Rachel M., Zhu, Zijie, Sun, Yaohui, Zhu, Kan, Xu, Zhengping, Lin, Francis, Pan, Tingrui, Losert, Wolfgang, Zhao, Min
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.05.2023; Elsevier
• Subjects: Biophysics; Biotechnology and bioengineering; Cell Biology; Cell-based Assays; Microscopy; Protocol; Tissue Engineering; Biophysics; Biotechnology and bioengineering; Microscopy; Cell-based Assays; Cell Biology; Tissue Engineering
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102288

Here, we present a protocol for electrotaxis of large epithelial cell sheets without compromising the integrity of cell epithelia in a high-throughput customized directed current electrotaxis chamber. We describe the fabrication and use of polydimethylsiloxane stencils to control the size and shape of human keratinocyte cell sheets. We detail cell tracking, cell sheet contour assay, and particle image velocimetry to reveal the spatial and temporal motility dynamics of cell sheets. This approach is applicable to other collective cell migration studies. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).1 [Display omitted] •An increased throughput system for electrotaxis of large epithelium cell sheets•Step-by-step guide for efficient patterning of epithelium cell sheets•Quantification of collective cell migration with PIV Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for electrotaxis of large epithelial cell sheets without compromising the integrity of cell epithelia in a high-throughput customized directed current electrotaxis chamber. We describe the fabrication and use of polydimethylsiloxane stencils to control the size and shape of human keratinocyte cell sheets. We detail cell tracking, cell sheet contour assay, and particle image velocimetry to reveal the spatial and temporal motility dynamics of cell sheets. This approach is applicable to other collective cell migration studies.