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Cells 2018, 7(5), 35; https://doi.org/10.3390/cells7050035

High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity

1
Research & Development Department, Cellular Technology Limited, Shaker Heights, OH 44122, USA
2
Institute of Anatomy and Cell Biology, Friedrich-Alexander University Erlangen-Nürnberg, 91054 Erlangen, Germany
*
Author to whom correspondence should be addressed.
Received: 27 March 2018 / Revised: 19 April 2018 / Accepted: 21 April 2018 / Published: 24 April 2018
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Abstract

One of the primary effector functions of immune cells is the killing of virus-infected or malignant cells in the body. Natural killer (NK) and CD8 effector T cells are specialized for this function. The gold standard for measuring such cell-mediated cytolysis has been the chromium release assay, in which the leakage of the radioactive isotope from damaged target cells is being detected. Flow cytometry-based single cell analysis of target cells has recently been established as a non-radioactive alternative. Here we introduce a target cell visualization assay (TVA) that applies similar target cell staining approaches as used in flow cytometry but based on single cell computer image analysis. Two versions of TVA are described here. In one, the decrease in numbers of calcein-stained, i.e., viable, target cells is assessed. In the other, the CFSE/PI TVA, the increase in numbers of dead target cells is established in addition. TVA assays are shown to operate with the same sensitivity as standard chromium release assays, and, leaving data audit trails in form of scanned (raw), analyzed, and quality-controlled images, thus meeting requirements for measuring cell-mediated cytolysis in a regulated environment. View Full-Text
Keywords: TVA; NK measurement; ADCC; CD8 cells; multi-color single cell imaging; CFR-Part 11 compliance; audit trails; cellular assay validation; immune monitoring TVA; NK measurement; ADCC; CD8 cells; multi-color single cell imaging; CFR-Part 11 compliance; audit trails; cellular assay validation; immune monitoring
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Welter, A.; Sundararaman, S.; Li, R.; Zhang, T.; Karulin, A.Y.; Lehmann, A.; Naeem, V.; Roen, D.R.; Kuerten, S.; Lehmann, P.V. High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring Cell-Mediated Cytotoxicity. Cells 2018, 7, 35.

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