A Novel Multiparametric Drug-Scoring Method for High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo Image Cytometer

Abstract

Three-dimensional (3D) tumor models have been increasingly used to investigate and characterize cancer drug compounds. The ability to perform high-throughput screening of 3D multicellular tumor spheroids (MCTS) can highly improve the efficiency and cost-effectiveness of discovering potential cancer drug candidates. Previously, the Celigo Image Cytometer has demonstrated a novel method for high-throughput screening of 3D multicellular tumor spheroids. In this work, we employed the Celigo Image Cytometer to examine the effects of 14 cancer drug compounds on 3D MCTS of the glioblastoma cell line U87MG in 384-well plates. Using parameters such as MCTS diameter and invasion area, growth and invasion were monitored for 9 and 3 d, respectively. Furthermore, fluorescent staining with calcein AM, propidium iodide, Hoechst 33342, and caspase 3/7 was performed at day 9 posttreatment to measure viability and apoptosis. Using the kinetic and endpoint data generated, we created a novel multiparametric drug-scoring system for 3D MCTS that can be used to identify and classify potential drug candidates earlier in the drug discovery process. Furthermore, the combination of quantitative and qualitative image data can be used to delineate differences between drugs that induce cytotoxic and cytostatic effects. The 3D MCTS-based multiparametric scoring method described here can provide an alternative screening method to better qualify tested drug compounds.

Real-Time Apoptosis and Viability High-Throughput Screening of 3D Multicellular Tumor Spheroids Using the Celigo Image Cytometer

“…The kinetic apoptosis and viability readouts in combination with bright-field growth inhibition data produce a unique multiplex orthogonal screen that can be used to evaluate hits but also elucidate potential mechanisms of action of these candidate drugs. 5 This multiplex screening approach can be used to further improve the quality and efficiency of the screen, resulting in identifying a more qualified drug candidate for drug discovery research.…”

Real‐time viability and apoptosis kinetic detection method of 3D multicellular tumor spheroids using the Celigo Image Cytometer

“…Future work will be to apply the proposed kinetic real-time viability and apoptosis detection method for MCTS to screen and characterize different drug compounds, and compare the results to the end point assay (9). By utilizing qualitative and quantitative kinetic data collected, it allows the researchers to properly establish a proper 3D MCTS model, determine when to treat the spheroids with test drugs, as well as characterization of drug effects.…”

“…As Celigo Image Cytometry can rapidly acquire and analyze spheroid images in bright field and fluorescence, we hypothesize that an initial primary drug screen would be conducted by performing end point-based assays, then a secondary drug screen would be conducted on the identified drug candidates by performing kinetic-based assays to determine the cytostatic, cytotoxic, and apoptotic effects to characterize target drug compounds. Future work will be to apply the proposed kinetic real-time viability and apoptosis detection method for MCTS to screen and characterize different drug compounds, and compare the results to the end point assay (9).…”

A rapid single cell sorting verification method using plate‐based image cytometry

“…Previously, the Celigo Image Cytometer has been utilized to perform cell‐based assays in microplates by capturing and analyzing whole well images . In this work, the Celigo Image Cytometer was used to rapidly count sorted single cells in Greiner 96‐well half area plates in order to determine the sorting efficiency of the FACS.…”