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Modular droplet-based microfluidic platform for functional phenotypic screening of natural killer cells

Natural Killer (NK) cells, as key effector cells of the innate immune system, display high heterogeneity in their ability to kill target cells. The underlying mechanisms remain poorly understood. Here, a droplet-based microfluidic platform is presented to identify and select NK cells with serial kil...

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Bibliographic Details
Main Authors: Aubermann, Florian (Author) , Seneca, Senne (Author) , Hofman, Tomáš (Author) , Garcés Lázaro, Irene (Author) , Ajmail, Karim (Author) , Daubner, Kai (Author) , Cerwenka, Adelheid (Author) , Platzman, Ilia (Author) , Spatz, Joachim P. (Author)
Format: Article (Journal)
Language:English
Published: 2025
In: Small Methods
Year: 2025, Pages: 1-9
ISSN:2366-9608
DOI:10.1002/smtd.202500236
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.1002/smtd.202500236
Verlag, kostenfrei, Volltext: http://onlinelibrary.wiley.com/doi/abs/10.1002/smtd.202500236
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Author Notes:Florian Aubermann, Senne Seneca, Tomáš Hofman, Irene Garcés-Lázaro, Karim Ajmail, Kai Daubner, Adelheid Cerwenka, Ilia Platzman, and Joachim P. Spatz
Description
Summary:Natural Killer (NK) cells, as key effector cells of the innate immune system, display high heterogeneity in their ability to kill target cells. The underlying mechanisms remain poorly understood. Here, a droplet-based microfluidic platform is presented to identify and select NK cells with serial killing ability. To this end, primary human NK cells are encapsulated with several target cells using an efficient negative pressure-based droplet generator. Capitalizing on the large number of possible killing events due to quantization into droplets, a convolutional neural network analysis pipeline is developed to quantify the cytotoxicity and abundance of serial killing events with high accuracy. To physically select NK cells based on their serial killing ability, MultiCell-Sort - an advanced real-time image-based droplet sorting module - is presented. While conventional single-cell sorters mostly evaluate intrinsically-encoded properties, such as protein expression levels, MultiCell-Sort can select living NK cells based on complex functional phenotypes emerging from multiple cell-cell interactions within the droplet. This novel microfluidic phenotyping platform hereby allows the potential integration of complementary techniques to provide an understanding of regulators and markers underlying the heterogeneous nature of NK cell functional phenotypes.
Item Description:Erstmals veröffentlicht: 21. April 2025
Gesehen am 27.05.2025
Physical Description:Online Resource
ISSN:2366-9608
DOI:10.1002/smtd.202500236

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