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

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.

SUBMITTER: Schwietzer MF

PROVIDER: S-EPMC10060904 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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

Schwietzer Mariel Flavia MF Thölmann Sonja S Greune Lilo L Ebnet Klaus K

STAR protocols 20230322 2

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 confoca ...[more]

PMID: 36952336

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