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

J Biomol Screen. 2016 Oct;21(9):931-41. doi: 10.1177/1087057116650965. Epub 2016 May 27.

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.

Keywords: assay validation; cell-based assays; oncology; patient-derived organoid culture.

MeSH terms

  • Antineoplastic Agents / isolation & purification*
  • Antineoplastic Agents / therapeutic use
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Colonic Neoplasms / drug therapy
  • Drug Discovery / methods
  • Drug Screening Assays, Antitumor / methods
  • High-Throughput Screening Assays / methods*
  • Humans
  • Organoids / growth & development*
  • Organoids / pathology
  • Spheroids, Cellular / drug effects

Substances

  • Antineoplastic Agents