A ring barrier-based migration assay to assess cell migration in vitro

Nature Protocols

Volumn 10, Issue 6, 2015, Pages 904-915

  Das, Asha M ^a   Eggermont, Alexander M M ^a,b   Ten Hagen, Timo L M ^a  

^a ERASMUS MEDICAL CENTER   (Netherlands)

^b INSTITUT GUSTAVE ROUSSY   (France)

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; AUTOMATION; BIOLOGICAL MONITORING; CELL INVASION; CELL MIGRATION; CELL MIGRATION ASSAY; CELLULAR PARAMETERS; CLINICAL PROTOCOL; COMPUTER ANALYSIS; CONTROLLED STUDY; CULTURE TECHNIQUE; EXTRACELLULAR MATRIX; HIGH THROUGHPUT SCREENING; HUMAN; HUMAN CELL; HUMAN CELL CULTURE; IN VITRO STUDY; MONOLAYER CULTURE; POLARIZATION; PRIORITY JOURNAL; REPRODUCIBILITY; RING BARRIER BASED MIGRATION ASSAY; SURFACE PROPERTY; CELL CULTURE; DEVICES; PROCEDURES;

CELL MIGRATION ASSAYS; CELLS, CULTURED; HUMANS;

EID: 84930069789     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2015.056     Document Type: Article

References (25)

1. Lauffenburger, D.A. & Horwitz, A.F. Cell migration: a physically integrated molecular process. Cell 84, 359-369 (1996).
2. Horwitz, A.R. & Parsons, J.T. Cell migration-movin' on. Science 286, 1102-1103 (1999).
3. Ridley, A.J. et al. Cell migration: integrating signals from front to back. Science 302, 1704-1709 (2003).
4. Petrie, R.J., Doyle, A.D. & Yamada, K.M. Random versus directionally persistent cell migration. Nat. Rev. Mol. Cell Biol. 10, 538-549 (2009).
5. Trinkaus, J.P. Directional cell movement during early development of the teleost Blennius pholis: I. Formation of epithelial cell clusters and their pattern and mechanism of movement. J. Exp. Zool 245, 157-186 (1988).
6. Fink, R.D. & Trinkaus, J.P. Fundulus deep cells: directional migration in response to epithelial wounding. Dev. Biol. 129, 179-190 (1988).
7. Cory, G. Scratch-wound assay. Methods Mol. Biol. 769, 25-30 (2011).
8. Liang, C.C., Park, A.Y. & Guan, J.L. in vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat. Protoc. 2, 329-333 (2007).
9. Qiang, L. et al. Regulation of cell proliferation and migration by p62 through stabilization of Twist1. Proc. Natl. Acad. Sci. USA 111, 9241-9246 (2014).
10. Varani, J., Orr, W. & Ward, P.A. A comparison of the migration patterns of normal and malignant cells in two assay systems. Am. J. Pathol. 90, 159-172 (1978).
11. Pratt, B.M., Harris, A.S., Morrow, J.S. & Madri, J.A. Mechanisms of cytoskeletal regulation. Modulation of aortic endothelial cell spectrin by the extracellular matrix. Am. J. Pathol. 117, 349-354 (1984).
12. Ohtaka, K., Watanabe, S., Iwazaki, R., Hirose, M. & Sato, N. Role of extracellular matrix on colonic cancer cell migration and proliferation. Biochem. Biophys. Res. Commun. 220, 346-352 (1996).
13. Van Horssen, R. & ten Hagen, T.L. Crossing barriers: the new dimension of 2D cell migration assays. J. Cell Physiol. 226, 288-290 (2011).
14. van Horssen, R., Galjart, N., Rens, J.A., Eggermont, A.M. & ten Hagen, T.L. Differential effects of matrix and growth factors on endothelial and fibroblast motility: application of a modified cell migration assay. J. Cell Biochem. 99, 1536-1552 (2006).
15. Bakker, E.R. et al. Wnt5a promotes human colon cancer cell migration and invasion but does not augment intestinal tumorigenesis in Apc1638N mice. Carcinogenesis 34, 2629-2638 (2013).
16. Das, A.M. et al. Differential TIMP3 expression affects tumor progression and angiogenesis in melanomas through regulation of directionally persistent endothelial cell migration. Angiogenesis 17, 163-177 (2014).
17. Cheng, C. et al. PDGF-induced migration of vascular smooth muscle cells is inhibited by heme oxygenase-1 via VEGFR2 upregulation and subsequent assembly of inactive VEGFR2/PDGFRβ heterodimers. Arterioscler. Thromb. Vasc. Biol. 32, 1289-1298 (2012).
18. van Horssen, R. et al. E-cadherin promotor methylation and mutation are inversely related to motility capacity of breast cancer cells. Breast Cancer Res. Treat. 136, 365-377 (2012).
19. Drabek, K. et al. Role of CLASP2 in microtubule stabilization and the regulation of persistent motility. Curr. Biol. 16, 2259-2264 (2006).
20. van Horssen, R. et al. Modulation of cell motility by spatial repositioning of enzymatic ATP/ADP exchange capacity. J. Biol. Chem. 284, 1620-1627 (2009).
21. Kuiper, J.W. et al. Local ATP generation by brain-type creatine kinase (CK-B) facilitates cell motility. PLoS ONE 4, e5030 (2009).
22. Gorelik, R. & Gautreau, A. Quantitative and unbiased analysis of directional persistence in cell migration. Nat. Protoc. 9, 1931-1943 (2014).
23. Deforet, M. et al. Automated velocity mapping of migrating cell populations (AVeMap). Nat. Methods 9, 1081-1083 (2012).
24. Zaritsky, A. et al. Cell motility dynamics: a novel segmentation algorithm to quantify multi-cellular bright-field microscopy images. PLoS ONE 6, e27593 (2011).
25. Huth, J. et al. Significantly improved precision of cell migration analysis in time-lapse video microscopy through use of a fully automated tracking system. BMC Cell Biol. 11, 24 (2010).