Abstract
An in vitro model for natural killer (NK) cell-mediated cytotoxicity was developed using a collagen-based scaffold 3D cell culture technology. This technology induced HCT116 cells to aggregate into tumoroids over time, which became the target cells during the cytotoxicity assay. Cytotoxicity was assessed by both phosphatidyl serine exposure (apoptosis) and plasma membrane rupture (necrosis) using fully automated workflows. Cytotoxicity was quantified using NK cells alone and with IL-2 stimulation, where a significant increase of cytotoxicity was evident. Cytotoxicity with this model was compared to HCT116 cells adhered to microplates in a conventional two-dimensional (2D) format. The three-dimensional (3D) model was far superior in both maintaining cell health over time and accurately depicting cytotoxic events.
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
Ohtani H (2007) Focus on TILs: prognostic significance of tumor infiltrating lymphocytes in human colorectal cancer. Cancer Immun 7(4):1–9
Bruno A, Ferlazzo G, Albini A, Noonan DM (2014) A think tank of TINK/TANKs: tumor-infiltrating/tumor-associated natural killer cells in tumor progression and angiogenesis. J Natl Cancer Inst 106(8):1–13
Eckl J, Buchner A, Prinz PU, Riesenberg R, Siegert SI, Kammerer R, Nelson PJ, Noessner E (2012) Transcript signature predicts tissue NK cell content and defines renal cell carcinoma subgroups independent of TNM staging. J Mol Med (Berl) 90(1):55–66
Hirschhaeuser F, Menne H, Dittfeld C, West J, Mueller-Klieser W, Kunz-Schughart LA (2010) Multicellular tumor tumoroids: an underestimated tool is catching up again. J Biotechnol 148(1):3–15
Blom WM, De Bont HJ, Meijerman I, Kuppen PJ, Mulder GJ, Nagelkerke JF (1999) Interleukin-2-activated natural killer cells can induce both apoptosis and necrosis in rat hepatocytes. Hepatology 29(3):785–792
Lehmann C, Zeis M, Uharek L (2001) Activation of natural killer cells with interleukin 2 (IL-2) and IL-12 increases perforin binding and subsequent lysis of tumour cells. Br J Haematol 114(3):660–665
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Larson, B., Hussain, L., Schroeder, J. (2020). Validation of an Image-Based 3D Natural Killer Cell-Mediated Cytotoxicity Assay. In: Tan, SL. (eds) Immuno-Oncology. Methods in Pharmacology and Toxicology. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0171-6_7
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0171-6_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0170-9
Online ISBN: 978-1-0716-0171-6
eBook Packages: Springer Protocols