A collagen-based microwell migration assay to study NK-target cell interactions.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
23 07 2019
23 07 2019
Historique:
received:
25
05
2018
accepted:
18
06
2019
entrez:
25
7
2019
pubmed:
25
7
2019
medline:
24
10
2020
Statut:
epublish
Résumé
Natural killer (NK) cell cytotoxicity in tissue is dependent on the ability of NK cells to migrate through the extracellular matrix (ECM) microenvironment. Traditional imaging studies of NK cell migration and cytotoxicity have utilized 2D surfaces, which do not properly reproduce the structural and mechanical cues that shape the migratory response of NK cells in vivo. Here, we have combined a microwell assay that allows long-term imaging and tracking of small, well-defined populations of NK cells with an interstitial ECM-like matrix. The assay allows for long-term imaging of NK-target cell interactions within a confined 3D volume. We found marked differences in motility between individual cells with a small fraction of the cells moving slowly and being confined to a small volume within the matrix, while other cells moved more freely. A majority of NK cells also exhibited transient variation in their motility, alternating between periods of migration arrest and movement. The assay could be used as a complement to in vivo imaging to study human NK cell heterogeneity in migration and cytotoxicity.
Identifiants
pubmed: 31337806
doi: 10.1038/s41598-019-46958-3
pii: 10.1038/s41598-019-46958-3
pmc: PMC6650390
doi:
Substances chimiques
Collagen
9007-34-5
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
10672Références
Front Immunol. 2014 Mar 07;5:80
pubmed: 24639676
Immunity. 2010 Oct 29;33(4):632-44
pubmed: 20951068
Nat Rev Immunol. 2006 Jul;6(7):497-507
pubmed: 16799470
Mol Biol Cell. 2017 Dec 1;28(25):3573-3581
pubmed: 29021341
Front Immunol. 2012 Nov 09;3:335
pubmed: 23162553
J Exp Med. 1999 Oct 4;190(7):1005-12
pubmed: 10510090
J Cell Biol. 1982 Mar;92(3):747-52
pubmed: 7085756
Immunity. 2006 Jul;25(1):129-41
pubmed: 16860762
Tissue Eng Part A. 2011 Mar;17(5-6):713-24
pubmed: 20929283
Curr Protoc Cell Biol. 2010 Sep;Chapter 10:Unit 10.18.1-20
pubmed: 20853341
Blood. 2013 Feb 21;121(8):1326-34
pubmed: 23287857
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15062-7
pubmed: 10611338
J Clin Invest. 2009 Jun;119(6):1420-8
pubmed: 19487818
Immunol Today. 1990 Jul;11(7):237-44
pubmed: 2201309
J Cell Biol. 1983 Apr;96(4):1089-96
pubmed: 6833393
Integr Biol (Camb). 2011 Jul;3(7):770-8
pubmed: 21687858
Biophys J. 2013 Nov 5;105(9):1967-75
pubmed: 24209841
Methods Mol Biol. 2016;1441:87-106
pubmed: 27177659
IEEE Trans Med Imaging. 2015 Apr;34(4):911-29
pubmed: 25415983
Nat Rev Immunol. 2009 Nov;9(11):789-98
pubmed: 19834485
Science. 2012 Jun 29;336(6089):1676-81
pubmed: 22745423
Elife. 2019 Sep 25;8:
pubmed: 31552826
Biomed Microdevices. 2011 Aug;13(4):683-93
pubmed: 21465090
Immunol Rev. 2013 Jan;251(1):154-9
pubmed: 23278747
Bioinformatics. 2014 Jun 1;30(11):1609-17
pubmed: 24526711
Cell Mol Life Sci. 2000 Jan 20;57(1):41-64
pubmed: 10949580
Microsc Res Tech. 1998 Dec 1;43(5):369-78
pubmed: 9858334
J Vis Exp. 2014 Oct 05;(92):e51963
pubmed: 25350138
PLoS Biol. 2005 Jun;3(6):e150
pubmed: 15857154
J Immunol. 2015 Oct 1;195(7):3374-81
pubmed: 26320254
Cold Spring Harb Perspect Biol. 2012 Jan 01;4(1):a004903
pubmed: 21937732
Biochem Pharmacol. 2010 Dec 15;80(12):2087-91
pubmed: 20849829
Cold Spring Harb Protoc. 2013 Feb 01;2013(2):100-17
pubmed: 23378652
Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):12081-6
pubmed: 17609379
Biophys J. 2010 Feb 3;98(3):L1-3
pubmed: 20141747
J Immunol. 2010 Mar 1;184(5):2329-36
pubmed: 20139277