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

Christopher J Bergin; Yannick D Benoit

doi:10.1016/j.xpro.2022.101218

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. eCollection 2022 Mar 18.

Authors

Christopher J Bergin¹, Yannick D Benoit¹

Affiliation

¹ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Colorectal Neoplasms* / pathology
Humans
Neoplastic Stem Cells / pathology
Organoids*