title: Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D.
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. First, spheroid arrays of 11 cell-lines were rapidly assessed for differences in spheroid morphology. Second, highly-positive (SOX-2), moderately-positive (Ki-67) and weakly-positive (βIII-tubulin) protein targets were detected and quantified. Third, the arrays enabled screening of ten media compositions for inducing differentiation in human neurospheres. Last, 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.
authors.affiliation: Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, Queen's Medical Centre, University of Nottingham, Nottingham, UK.
authors.forename: Delyan P
authors.initials: DP
authors.lastname: Ivanov
authors.affiliation: Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, Queen's Medical Centre, University of Nottingham, Nottingham, UK.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Grabowska
mesh_terms: D000465:Algorithms; D015415:Biomarkers / Q000378:metabolism*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D045325:HCT116 Cells; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D019394:Ki-67 Antigen / Q000378:metabolism; D061986:MCF-7 Cells; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D018874:Spheroids, Cellular / Q000166:cytology*; D046888:Tissue Array Analysis / Q000295:instrumentation*; D014404:Tubulin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D019394:Ki-67 Antigen; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C498585:TUBB3 protein, human; D014404:Tubulin