Protocol for electrotaxis of large epithelial cell sheets

Yan Zhang; Rachel M Lee; Zijie Zhu; Yaohui Sun; Kan Zhu; Zhengping Xu; Francis Lin; Tingrui Pan; Wolfgang Losert; Min Zhao

doi:10.1016/j.xpro.2023.102288

Protocol for electrotaxis of large epithelial cell sheets

STAR Protoc. 2023 May 6;4(2):102288. doi: 10.1016/j.xpro.2023.102288. Online ahead of print.

Authors

Yan Zhang¹, Rachel M Lee², Zijie Zhu³, Yaohui Sun⁴, Kan Zhu⁴, Zhengping Xu⁵, Francis Lin⁶, Tingrui Pan⁷, Wolfgang Losert⁸, Min Zhao⁹

Affiliations

¹ Department of Ophthalmology & Vision Science, University of California, California, Davis, CA 95616, USA; School of Public Health, Hangzhou Normal University, Hangzhou 311121, China; Institute of Environmental Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China; Micro-Nano Innovations (MiNI) Laboratory, Department of Biomedical Engineering, University of California, California, Davis, CA 95616, USA. Electronic address: yanzhang@hznu.edu.cn.
² Institute for Physical Science and Technology, University of Maryland, College Park, MD 20742, USA.
³ Micro-Nano Innovations (MiNI) Laboratory, Department of Biomedical Engineering, University of California, California, Davis, CA 95616, USA.
⁴ Department of Ophthalmology & Vision Science, University of California, California, Davis, CA 95616, USA.
⁵ Institute of Environmental Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
⁶ Department of Physics and Astronomy, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
⁷ Micro-Nano Innovations (MiNI) Laboratory, Department of Biomedical Engineering, University of California, California, Davis, CA 95616, USA; Institute of Biomedical and Health Engineering, Shenzhen Institute of Advanced Technology, Shenzhen 518055, China; Shenzhen Engineering Laboratory of Single-molecule Detection and Instrument Development, Shenzhen, Guangdong 518055, China; Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou 215123, China.
⁸ Institute for Physical Science and Technology, University of Maryland, College Park, MD 20742, USA; Department of Physics, University of Maryland, College Park, MD 20742, USA.
⁹ Department of Ophthalmology & Vision Science, University of California, California, Davis, CA 95616, USA; Department of Dermatology, University of California, California, Davis, CA 95616, USA. Electronic address: minzhao@ucdavis.edu.

Abstract

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.

Keywords: Biophysics; Biotechnology and bioengineering; Cell Biology; Cell-based Assays; Microscopy; Tissue Engineering.