FACS-based protocol to assess cytotoxicity and clonogenic potential of colorectal cancer stem cells using a Wnt/β-catenin signaling pathway reporter

Simone Di Franco; Le Zhang; Miriam Gaggianesi; Melania Lo Iacono; Jan Paul Medema; Giorgio Stassi

STAR Protocols (Dec 2021)

FACS-based protocol to assess cytotoxicity and clonogenic potential of colorectal cancer stem cells using a Wnt/β-catenin signaling pathway reporter

Simone Di Franco,
Le Zhang,
Miriam Gaggianesi,
Melania Lo Iacono,
Jan Paul Medema,
Giorgio Stassi

Affiliations

Simone Di Franco: Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy
Le Zhang: Laboratory for Experimental Oncology and Radiobiology, Center for Experimental and Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands; Oncode Institute, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands
Miriam Gaggianesi: Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy
Melania Lo Iacono: Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy
Jan Paul Medema: Laboratory for Experimental Oncology and Radiobiology, Center for Experimental and Molecular Medicine, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands; Oncode Institute, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands
Giorgio Stassi: Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 100880

Abstract

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Summary: Cancer stem cells (CSCs) play a key role in tumor initiation and progression. A real-time tool to evaluate the activation of CSC-specific signaling pathways is crucial for the study of this cancer cell subset. Here, we present a protocol to monitor, in vitro, the activation of Wnt/β-catenin signaling pathway, which is considered a functional biomarker for colorectal CSCs (CR-CSCs). This flow-cytometry-based protocol allows it to isolate CR-CSCs and to evaluate their cytotoxicity upon anti-tumor treatments.For complete details on the use and execution of this protocol, please refer to Di Franco et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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