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  Droplet‐based combinatorial assay for cell cytotoxicity and cytokine release evaluation

Antona, S., Abele, T., Jahnke, K., Dreher, Y., Göpfrich, K., Platzman, I., et al. (2020). Droplet‐based combinatorial assay for cell cytotoxicity and cytokine release evaluation. Advanced Functional Materials, 30(46): 2003479, pp. 1-9. doi:10.1002/adfm.202003479.

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 Creators:
Antona, Silvia1, Author              
Abele, Tobias1, Author              
Jahnke, Kevin1, Author              
Dreher, Yannik1, Author              
Göpfrich, Kerstin1, Author              
Platzman, Ilia1, Author              
Spatz, Joachim P.1, Author              
Affiliations:
1Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_2364731              

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Free keywords: cytotoxicity assay; droplet‐based microfluidics; interferon‐gamma; natural killer cells; single‐cell analysis
 Abstract: Balancing the maximal efficacy of interferon gamma (IFN‐ɣ)‐based therapies with its side effects is a great challenge for future cytokine treatments. To achieve this, the development of single‐cell technologies that study IFN‐ɣ release in correlation with antitumor activity would represent a huge step forward. To this end, droplet‐based microfluidics is employed to quantitatively investigate IFN‐ɣ secretion from single natural killer (NK) cells in correlation with their cytotoxic activity against a specific target. The method relies on co‐encapsulation of NK‐92 cells, target cancer cells, polystyrene beads conjugated with specific IFN‐ɣ capture antibodies, and fluorescently labeled detection antibodies inside water‐in‐oil compartments. The secreted cytokines are captured and detected by localized fluorescence at the periphery of the beads. NK‐92's cytotoxicity is evaluated simultaneously by means of a fluorescent DNA intercalating agent, which penetrates the membranes of dead target cells. To deepen the understanding of the role of the cytokine in antitumor immunomodulation, the impact of different doses of human recombinant IFN‐ɣ on the cytolytic activity of NK‐92 cells shows a trend that the higher the dose the lower the cytolytic activity of NK cells. The developed method represents a simple quantitative approach to unravel the complex heterogeneity of NK cells toward IFN‐ɣ secretion and cytolytic activity.

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Language(s): eng - English
 Dates: 2020-08-092020-04-202020-09-202020-11-11
 Publication Status: Published in print
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/adfm.202003479
 Degree: -

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Title: Advanced Functional Materials
  Other : Adv. Funct. Mater.
Source Genre: Journal
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Publ. Info: Weinheim : Wiley-VCH Verlag GmbH
Pages: - Volume / Issue: 30 (46) Sequence Number: 2003479 Start / End Page: 1 - 9 Identifier: ISSN: 1616-301X
CoNE: https://pure.mpg.de/cone/journals/resource/954925596563