High-content assays for characterizing the viability and morphology of 3D cancer spheroid cultures

Assay and Drug Development Technologies
Oksana SirenkoEvan F Cromwell

Abstract

There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translation research. Development of higher throughput assays to quantify phenotypic changes in spheroids is an active area of investigation. The goal of this study was to develop higher throughput high-content imaging and analysis methods to characterize phenotypic changes in human cancer spheroids in response to compound treatment. We optimized spheroid cell culture protocols using low adhesion U-bottom 96- and 384-well plates for three common cancer cell lines and improved the workflow with a one-step staining procedure that reduces assay time and minimizes variability. We streamlined imaging acquisition by using a maximum projection algorithm that combines cellular information from multiple slices through a 3D object into a single image, enabling efficient comparison of different spheroid phenotypes. A custom image analysis method was implemented to provide multiparametric characterization of single-cell and spheroid phenotypes. We report a number of readouts, including quantification of marker-specific cell numbers, measurement of cell viability and apoptosis, and characterization of spheroid size ...Continue Reading

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Methods Mentioned

BETA
imaging methods
Assay
light scattering

Software Mentioned

MetaXpress CME
MetaXpress Module Editor
MaxPro
SoftMax ® Pro
Cell Scoring
CellEvent
MetaXpress

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