Advertisement

Random Migration Assays of Mammalian Cells and Quantitative Analyses of Single Cell Trajectories

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1749)

Abstract

Cell migration is essential to many biological processes such as embryonic development, immune surveillance and wound healing. Random cell migration refers to the intrinsic ability of cells to migrate, often called cell motility. This basal condition contrasts with directed cell migration, where cells migrate toward a chemical or physical cue. Unlike Brownian particles, however, randomly migrating cells exhibit a directional persistence, i.e., they are more likely to sustain the movement in the direction they previously took than to change, even if this direction is randomly chosen in an isotropic environment. Here we describe how to set up time-lapse recording of mammalian cells freely moving on a two-dimensional surface coated with extracellular matrix proteins, how to acquire single cell trajectories from movies and how to extract key parameters that characterize cell motility, such as cell speed, directionality, mean square displacement, and directional persistence.

Key words

Cell tracking Speed Directionality MSD Persistence Autocorrelation 

References

  1. 1.
    Sanz-Moreno V, Gadea G, Ahn J et al (2008) Rac activation and inactivation control plasticity of tumor cell movement. Cell 135:510–523.  https://doi.org/10.1016/j.cell.2008.09.043 CrossRefPubMedGoogle Scholar
  2. 2.
    Sanz-Moreno V, Marshall CJ (2010) The plasticity of cytoskeletal dynamics underlying neoplastic cell migration. Curr Opin Cell Biol 22:690–696.  https://doi.org/10.1016/j.ceb.2010.08.020 CrossRefPubMedGoogle Scholar
  3. 3.
    Liu YJ, Le Berre M, Lautenschlaeger F et al (2015) Confinement and low adhesion induce fast amoeboid migration of slow mesenchymal cells. Cell 160:659–672.  https://doi.org/10.1016/j.cell.2015.01.007 CrossRefPubMedGoogle Scholar
  4. 4.
    Panopoulos A, Howell M, Fotedar R, Margolis RL (2011) Glioblastoma motility occurs in the absence of actin polymer. Mol Biol Cell 22:2212–2220.  https://doi.org/10.1091/mbc.E10-10-0849 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Dang I, Gorelik R, Sousa-Blin C et al (2013) Inhibitory signalling to the Arp2/3 complex steers cell migration. Nature 503:281–284.  https://doi.org/10.1038/nature12611 CrossRefPubMedGoogle Scholar
  6. 6.
    Gorelik R, Gautreau A (2015) The Arp2/3 inhibitory protein arpin induces cell turning by pausing cell migration. Cytoskeleton 72:362–371.  https://doi.org/10.1002/cm.21233 CrossRefPubMedGoogle Scholar
  7. 7.
    Petrie RJ, Doyle AD, Yamada KM (2009) Random versus directionally persistent cell migration. Nat Rev Mol Cell Biol 10:538–549.  https://doi.org/10.1038/nrm2729 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Krause M, Gautreau A (2014) Steering cell migration: lamellipodium dynamics and the regulation of directional persistence. Nat Rev Mol Cell Biol 15:577–590.  https://doi.org/10.1038/nrm3861 CrossRefPubMedGoogle Scholar
  9. 9.
    Meijering E, Dzyubachyk O, Smal I (2012) Methods for cell and particle tracking. Methods Enzymol 504:183–200.  https://doi.org/10.1016/B978-0-12-391857-4.00009-4 CrossRefPubMedGoogle Scholar
  10. 10.
    Gorelik R, Gautreau A (2014) Quantitative and unbiased analysis of directional persistence in cell migration. Nat Protoc 9:1931–1943.  https://doi.org/10.1038/nprot.2014.131 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Department of Systems Pharmacology and Translational TherapeuticsUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Ecole Polytechnique, Université Paris-Saclay, BIOC-CNRS UMR7654PalaiseauFrance
  3. 3.School of Biological and Medical PhysicsMoscow Institute of Physics and TechnologyMoscow RegionRussian Federation

Personalised recommendations