Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures. (October 2016)

• Record Type: Journal Article
• Authors: Boehnke, Karsten; Iversen, Philip W.; Schumacher, Dirk; Lallena, María José; Haro, Rubén; Amat, Joaquín; Haybaeck, Johannes; Liebs, Sandra; Lange, Martin; Schäfer, Reinhold; Regenbrecht, Christian R. A.; Reinhard, Christoph; Velasco, Juan A.
• Is Part Of: SLAS discovery. Volume 21:Number 9(2016)
• Page Start: 931
• Page End: 941
• Publication Date: 2016-10
• Subjects: cell-based assays -- oncology -- assay validation -- patient-derived organoid culture; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; Biomolecules -- Analysis; Drugs -- Analysis; Drugs -- Testing; Drug Evaluation, Preclinical; Molecular Biology -- methods; Periodicals; Periodicals; 615.1
• DOI: 10.1177/1087057116650965
• Languages: English
• ISSNs: 2472-5552

Abstract

The application of patient-derived three-dimensional culture systems as disease-specific drug sensitivity models has enormous potential to connect compound screening and clinical trials. However, the implementation of complex cell-based assay systems in drug discovery requires reliable and robust screening platforms. Here we describe the establishment of an automated platform in 384-well format for three-dimensional organoid cultures derived from colon cancer patients. Single cells were embedded in an extracellular matrix by an automated workflow and subsequently self-organized into organoid structures within 4 days of culture before being exposed to compound treatment. We performed validation of assay robustness and reproducibility via plate uniformity and replicate-experiment studies. After assay optimization, the patient-derived organoid platform passed all relevant validation criteria. In addition, we introduced a streamlined plate uniformity study to evaluate patient-derived colon cancer samples from different donors. Our results demonstrate the feasibility of using patient-derived tumor samples for high-throughput assays and their integration as disease-specific models in drug discovery.