A micropattern-based assay to study contact inhibition of locomotion and entosis of adherent human and canine cells in vitro

Mariel Flavia Schwietzer; Sonja Thölmann; Lilo Greune; Klaus Ebnet

doi:10.1016/j.xpro.2023.102186

A micropattern-based assay to study contact inhibition of locomotion and entosis of adherent human and canine cells in vitro

STAR Protoc. 2023 Mar 22;4(2):102186. doi: 10.1016/j.xpro.2023.102186. Online ahead of print.

Authors

Mariel Flavia Schwietzer¹, Sonja Thölmann², Lilo Greune³, Klaus Ebnet⁴

Affiliations

¹ Institute-associated Research Group "Cell Adhesion and Cell Polarity", Institute of Medical Biochemistry, ZMBE, University of Münster, 48149 Münster, Germany. Electronic address: [email protected].
² Institute-associated Research Group "Cell Adhesion and Cell Polarity", Institute of Medical Biochemistry, ZMBE, University of Münster, 48149 Münster, Germany.
³ Institute of Infectiology, ZMBE, University of Münster, 48149 Münster, Germany.
⁴ Institute-associated Research Group "Cell Adhesion and Cell Polarity", Institute of Medical Biochemistry, ZMBE, University of Münster, 48149 Münster, Germany. Electronic address: [email protected].

Abstract

We present a protocol for using micropatterns to study post-collision locomotion and entosis of human and canine cells in vitro. We describe steps for lentiviral transduction and the preparation of micropatterned slides consisting of narrow matrix-coated stripes separated by cytophobic spacers. We then detail cell seeding, chamber assembly, and live cell analysis. We provide steps for analysis by live cell imaging using fluorescence microscopy as well as fixing for subsequent analysis by confocal microscopy or correlative light and electron microscopy. For complete details on the use and execution of this protocol, please refer to Kummer et al. (2022)¹ and Schwietzer et al. (2022).².

Keywords: Cell-based Assays; Single Cell.