Protocol to generate scaffold-free, multicomponent 3D melanoma spheroid models for preclinical drug testing

STAR Protoc. 2024 Jun 21;5(2):103058. doi: 10.1016/j.xpro.2024.103058. Epub 2024 May 14.

Abstract

Three-dimensional (3D) models play an increasingly important role in preclinical drug testing as they faithfully mimic interactions between cancer cells and the tumor microenvironment (TME). Here, we present a protocol for generating scaffold-free 3D multicomponent human melanoma spheroids. We describe steps for characterizing models using live-cell imaging and histology, followed by drug testing and assessment of cell death through various techniques such as imaging, luminescence-based assays, and flow cytometry. Finally, we demonstrate the models' adaptability for co-cultures with immune cells.

Keywords: Cancer; Cell Biology; High Throughput Screening; Organoids.

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Line, Tumor
  • Coculture Techniques / methods
  • Drug Evaluation, Preclinical / methods
  • Drug Screening Assays, Antitumor / methods
  • Humans
  • Melanoma* / metabolism
  • Melanoma* / pathology
  • Spheroids, Cellular* / metabolism
  • Spheroids, Cellular* / pathology
  • Tumor Microenvironment