Protocol for serial organoid formation assay using primary colorectal cancer tissues to evaluate cancer stem cell activity

  • STAR Protoc. 2022 Mar 4;3(1):101218. doi: 10.1016/j.xpro.2022.101218.
Christopher J Bergin  1 ,  Yannick D Benoit  1
Affiliations
  • 1. Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
PMID: 35265864 DOI: 10.1016/j.xpro.2022.101218
Abstract

Organoids can enable the study of solid tumors initiated from a single Cancer stem cell (CSC) ex vivo. We describe a serial tumor Organoid plating protocol using primary colorectal Cancer (CRC) tissues as a rapid and cost-efficient approach to evaluate the impact of therapeutic interventions on CSC functions. We detail the isolation of primary colorectal CSCs, Organoid embedding, serial passaging, and CSC-related analytical techniques. For complete details on the use and execution of this protocol, please refer to Masibag et al. (2021) and Bergin et al. (2021).

Keywords
Cancer; Cell isolation; Cell-based Assays; Health Sciences; Microscopy; Organoids; Single Cell; Stem Cells; Tissue Engineering.
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