Abstract
The rapid, efficient detection of cell death is critical for characterizing the underlying biology of in vitro disease models and, in particular, immunotherapy products used for preclinical therapeutic research. Traditional endpoint assays are laborious to perform for mass screening of therapeutic candidates and may fail to fully capture the kinetics of events surrounding the initiation, duration, and mechanisms of cell death—important events that may affect translational relevance and impact therapeutic decision-making during development. Here, we describe simple, efficient methods to measure apoptosis and immune cell killing in both adherent and nonadherent cell populations using the Incucyte® Live-Cell Analysis system and associated nonperturbing reagents, cells, and protocols. Assays are performed in the user’s own incubator with minimal disturbance and may be readily incorporated into existing workflows. Users may multiplex to maximize data collection from each sample. The integrated, user-friendly software does not require advanced technical training, enabling rapid analysis. Taken together, this method provides essential kinetic insight for greater understanding of cell death and the dynamic interactions between immune cells and their targets.
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References
Bernards R et al (2020) A roadmap for the next decade in cancer research. Nat Cancer 1:12–17
Roth TL et al (2018) Reprogramming human T cell function and specificity with non-viral genome targeting. Nature 559(7714):405–409
Shifrut E et al (2018) Genome-wide CRISPR screens in primary human T cell reveal key regulators of immune functions. Cell 175(7):1958–1971
Fry TJ et al (2018) CD22-targeted CAR T cells induce remission in B=ALL that is naive or resistant to CD19-targeted CAR immunotherapy. Nat Med 24(1):20–28
Further Reading
Additional protocols available online at EssenBioscience.com. Obtain a copy of the third edition, Live-cell analysis handbook: https://www.essenbioscience.com/en/communications/live-cell-analysis-handbook-4th-edition/
Acknowledgement
The science presented in this chapter is a product of the sustained, multidisciplinary R&D activities of Essen BioScience/Sartorius personnel based in Ann Arbor, Michigan, USA, and Welwyn Garden City, Hertfordshire, UK. This includes cell biologists, software engineers, optical and hardware designers as well as essential backup support functions, communications, and product development. We thank you for your hard work and dedication to move science forward. We also thank our customers for their feedback and creativity, which continues to inspire us.
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Granger, J.E., Appledorn, D.M. (2021). Kinetic Measurement of Apoptosis and Immune Cell Killing Using Live-Cell Imaging and Analysis. In: Alvero, A.B., Mor, G.G. (eds) Detection of Cell Death Mechanisms. Methods in Molecular Biology, vol 2255. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1162-3_17
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DOI: https://doi.org/10.1007/978-1-0716-1162-3_17
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1161-6
Online ISBN: 978-1-0716-1162-3
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