Abstract
Neutrophil migration from the bloodstream to sites of infection or injury is a multistep process that requires, dependent on the tissue structures being encountered, different modes of movement. Neutrophil locomotion can range from mesenchymal to amoeboid movement and may include multiple shape changes, contractile squeezing through gaps, and adhesion/de-adhesion cycles. In vitro migration assays reflect only some aspects of the complex in vivo neutrophil recruitment. For two-dimensional in vitro migration chemotaxis chambers, microscopic analysis of movement towards a pipette gradient or Boyden chambers is used. To analyze three-dimensional in vitro migration neutrophils can be embedded into matrices of diverse biophysical properties or can be placed onto matrices that are layered on a wide-pore filter, enabling migration through the matrix and the filter of a transwell plate towards a gradient of chemoattractant. We utilize here a commercially available setup for migration of murine neutrophils from the top of a loose collagen type I matrix, which determines the ability of neutrophils to attach to the matrix, sense the chemoattractant, polarize, digest the matrix, and move through the matrix into the lower transwell chamber. While the mode of migration inside the matrix cannot be studied in detail, this assay permits quantitative assessment of migrated neutrophils during a defined period of time.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Borregaard N (2010) Neutrophils, from marrow to microbes. Immunity 33:657–670
Butler KL, Ambravaneswaran V, Agrawal N et al (2010) Burn injury reduces neutrophil directional migration speed in microfluidic devices. PLoS ONE 5:e11921
Nourshargh S, Hordijk PL, Sixt M (2010) Breaching multiple barriers: leukocyte motility through vascular walls and the interstitium. Nat Rev Mol Cell Biol 11:366–378
Huttenlocher A, Horwitz AR (2011) Integrins in cell migration. Cold Spring Harb Perspect Biol 3:a005074
Lammermann T, Sixt M (2009) Mechanical modes of ‘amoeboid’ cell migration. Curr Opin Cell Biol 21:636–644
Yoshida K, Kondo R, Wang Q et al (2006) Neutrophil cytoskeletal rearrangements during capillary sequestration in bacterial pneumonia in rats. Am J Respir Crit Care Med 174: 689–698
Wolf K, Friedl P (2011) Extracellular matrix determinants of proteolytic and non-proteolytic cell migration. Trends Cell Biol 21:736–744
Van Goethem E, Poincloux R, Gauffre F et al (2010) Matrix architecture dictates three-dimensional migration modes of human macrophages: differential involvement of proteases and podosome-like structures. J Immunol 184: 1049–1061
Koenderman L, Van der Linden JAM, Honing H et al (2010) Integrins on neutrophils are dispensable for migration into three-dimensional fibrin gels. Thromb Haemost 104:599–608
Zaman MH, Trapani LM, Sieminski AL et al (2006) Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. Proc Natl Acad Sci U S A 103:10889–10894
Duong H, Wu B, Tawil B (2009) Modulation of 3D fibrin matrix stiffness by intrinsic fibrinogen–thrombin compositions and by extrinsic cellular activity. Tissue Eng A 15:1865–1876
Wang Q, Teder P, Judd NP et al (2002) CD44 deficiency leads to enhanced neutrophil migration and lung injury in Escherichia coli pneumonia in mice. Am J Pathol 161:2219–2228
Steadman R, St John PL, Evans RA et al (1997) Human neutrophils do not degrade major basement membrane components during chemotactic migration. Int J Biochem Cell Biol 29:993–1004
Sixt M, Lammermann T (2011) In vitro analysis of chemotactic leukocyte migration in 3D environments. Methods Mol Biol 769: 149–165
Acknowledgments
This work was supported by grants from Science Foundation Ireland and the Health Research Board.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Jennings, R.T., Knaus, U.G. (2014). Neutrophil Migration Through Extracellular Matrix. In: Quinn, M., DeLeo, F. (eds) Neutrophil Methods and Protocols. Methods in Molecular Biology, vol 1124. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-845-4_13
Download citation
DOI: https://doi.org/10.1007/978-1-62703-845-4_13
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-844-7
Online ISBN: 978-1-62703-845-4
eBook Packages: Springer Protocols