Abstract
Effector functions and cellular properties of natural killer (NK) cells are regulated by cellular and extracellular factors shaped by the microenvironments. NK cells express specific chemokine and non-chemokine receptors to aid preferential migrations or localizations in tissues. Good understanding of how NK-cell migratory properties are regulated in physiological and pathological microenvironments will provide further insights into the development of NK cell-based therapeutic approaches. In contrast to the commonly used conventional in vitro migration assays such as Trans-well assays that measure movements of a population of the migratory cells, microfluidic-based devices support live-cell imaging of cell migrations under a well-defined chemical gradient(s) at microscale. Subsequent analyses at single-cell level provide quantitative measurements of cell-migration parameters such as speed and Chemotactic Index, and permit distinguishing chemotaxis, chemokinesis, and chemo-repulsion. Our recent work established the use of a Y-shaped microfluidic device to study NK cell migrations in vitro. In this chapter, we described the detailed method of acquiring and analyzing NK cell migration in the microfluidic devices.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Horwitz R, Webb D (2003) Cell migration. Curr Biol 13:R756–R759
Friedl P, Weigelin B (2008) Interstitial leukocyte migration and immune function. Nat Immunol 9:960–969
Luster AD, Alon R, von Andrian UH (2005) Immune cell migration in inflammation: present and future therapeutic targets. Nat Immunol 6:1182–1190
Shi FD, Zhou Q (2011) Natural killer cells as indispensable players and therapeutic targets in autoimmunity. Autoimmunity 44:3–10
Orr MT, Lanier LL (2010) Natural killer cell education and tolerance. Cell 142:847–856
Waldhauer I, Steinle A (2008) NK cells and cancer immunosurveillance. Oncogene 27:5932–5943
Walzer T, Chiossone L, Chaix J et al (2007) Natural killer cell trafficking in vivo requires a dedicated sphingosine 1-phosphate receptor. Nat Immunol 8:1337–1344
Maghazachi AA (2010) Role of chemokines in the biology of natural killer cells. Curr Top Microbiol Immunol 341:37–58
Somanchi SS, Somanchi A, Cooper LJ et al (2012) Engineering lymph node homing of ex vivo-expanded human natural killer cells via trogocytosis of the chemokine receptor CCR7. Blood 119:5164–5172
Boyden S (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466
Li Jeon N, Baskaran H, Dertinger SK et al (2002) Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device. Nat Biotechnol 20:826–830
Chiu DT, Jeon NL, Huang S et al (2000) Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems. Proc Natl Acad Sci U S A 97:2408–2413
Lin F, Nguyen CM, Wang SJ et al (2005) Neutrophil migration in opposing chemoattractant gradients using microfluidic chemotaxis devices. Ann Biomed Eng 33:475–482
Lin F, Butcher EC (2006) T cell chemotaxis in a simple microfluidic device. Lab Chip 6:1462–1469
Mahmood S, Nandagopal S, Sow I et al (2014) Microfluidic-based, live-cell analysis allows assessment of NK-cell migration in response to crosstalk with dendritic cells. Eur J Immunol 44:2737–2748
Wang H, Zhao C, Zhang M et al (2013) A novel role of the scaffolding protein JLP in tuning CD40-induced activation of dendritic cells. Immunobiology 218:835–843
Chung BG, Lin F, Jeon NL (2006) A microfluidic multi-injector for gradient generation. Lab Chip 6:764–768
Nandagopal S, Wu D, Lin F (2011) Combinatorial guidance by CCR7 ligands for T lymphocytes migration in co-existing chemokine fields. PLoS One 6:e18183
Acknowledgement
This work was supported by Establishment and Operating Grants from the Manitoba Health Research Council, and Natural Sciences and Engineering Research Council (RGPIN/04775-2014 to S.K.P.K.). S.N. is supported by a postdoctoral fellowship provided by Terry Fox Foundation, Manitoba Institute of Child Health, and Research Manitoba, and in part by an operating grant from Canadian Cancer Society Research Institute (to S.K.P.K.). This work is supported by NSERC Discovery Grant (to S.K.P.K.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Nandagopal, S., Lin, F., Kung, S.K.P. (2016). Microfluidic-Based Live-Cell Analysis of NK Cell Migration In Vitro. In: Somanchi, S. (eds) Natural Killer Cells. Methods in Molecular Biology, vol 1441. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3684-7_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3684-7_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3682-3
Online ISBN: 978-1-4939-3684-7
eBook Packages: Springer Protocols