title: Translation of a tumor microenvironment mimicking 3D tumor growth co-culture assay platform to high-content screening.
abstract: For drug discovery, cell-based assays are becoming increasingly complex to mimic more realistically the nature of biological processes and their diversifications in diseases. Multicellular co-cultures embedded in a three-dimensional (3D) matrix have been explored in oncology to more closely approximate the physiology of the human tumor microenvironment. High-content analysis is the ideal technology to characterize these complex biological systems, although running such complex assays at higher throughput is a major endeavor. Here, we report on adapting a 3D tumor co-culture growth assay to automated microscopy, and we compare various imaging platforms (confocal vs. nonconfocal) with correlating automated image analysis solutions to identify optimal conditions and settings for future larger scaled screening campaigns. The optimized protocol has been validated in repeated runs where established anticancer drugs have been evaluated for performance in this innovative assay.
authors.affiliation: Janssen R&D, a Division of Janssen Pharmaceutica NV, Beerse, Belgium. ekrausz@its.jnj.com
authors.forename: Eberhard
authors.initials: E
authors.lastname: Krausz
authors.forename: Ronald
authors.initials: R
authors.lastname: de Hoogt
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Gustin
authors.forename: Frans
authors.initials: F
authors.lastname: Cornelissen
authors.forename: Thierry
authors.initials: T
authors.lastname: Grand-Perret
authors.forename: Lut
authors.initials: L
authors.lastname: Janssen
authors.forename: Nele
authors.initials: N
authors.lastname: Vloemans
authors.forename: Dirk
authors.initials: D
authors.lastname: Wuyts
authors.forename: Sandy
authors.initials: S
authors.lastname: Frans
authors.forename: Amy
authors.initials: A
authors.lastname: Axel
authors.forename: Pieter Johan
authors.initials: PJ
authors.lastname: Peeters
authors.forename: Brett
authors.initials: B
authors.lastname: Hall
authors.forename: Miroslav
authors.initials: M
authors.lastname: Cik
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004354:Drug Screening Assays, Antitumor / Q000379:methods* / Q000592:standards; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards; D006801:Humans; D007091:Image Processing, Computer-Assisted; D020128:Inhibitory Concentration 50; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects; D012015:Reference Standards; D012984:Software; D059016:Tumor Microenvironment
chemical_list: D000970:Antineoplastic Agents