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Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D

Ivanov, Delyan P.; Grabowska, Anna M.

Authors

Delyan P. Ivanov

ANNA GRABOWSKA ANNA.GRABOWSKA@NOTTINGHAM.AC.UK
Professor of Cancer Microenvironment



Abstract

We describe and share a device, methodology and image analysis algorithms, which allow up to 66 spheroids to be arranged into a gel-based array directly from a culture plate for downstream processing and analysis. Compared to processing individual samples, the technique uses 11-fold less reagents, saves time and enables automated imaging. To illustrate the power of the technology, we showcase applications of the methodology for investigating 3D spheroid morphology and marker expression and for in vitro safety and efficacy screens. Firstly, spheroid arrays of 11 cell-lines were rapidly assessed for differences in spheroid morphology. Secondly, highly-positive (SOX-2), moderately-positive (Ki-67) and weakly-positive (?III-tubulin) protein targets were detected and quantified. Third, the arrays enabled screening of ten media compositions for inducing differentiation in human neurospheres. Lastly, the application of spheroid microarrays for spheroid-based drug-screens was demonstrated by quantifying the dose-dependent drop in proliferation and increase in differentiation in etoposide-treated neurospheres.

Citation

Ivanov, D. P., & Grabowska, A. M. (2017). Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D. Scientific Reports, 7(1), Article 41160. https://doi.org/10.1038/srep41160

Journal Article Type Article
Acceptance Date Dec 14, 2016
Online Publication Date Jan 30, 2017
Publication Date 2017-12
Deposit Date Jan 17, 2017
Publicly Available Date Mar 29, 2024
Journal Scientific Reports
Electronic ISSN 2045-2322
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 7
Issue 1
Article Number 41160
DOI https://doi.org/10.1038/srep41160
Public URL https://nottingham-repository.worktribe.com/output/838657
Publisher URL http://www.nature.com/articles/srep41160

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