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

doi:10.1016/j.xpro.2021.100880

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

STAR Protoc. 2021 Oct 19;2(4):100880. doi: 10.1016/j.xpro.2021.100880. eCollection 2021 Dec 17.

Authors

Simone Di Franco¹, Le Zhang^{2

3}, Miriam Gaggianesi¹, Melania Lo Iacono¹, Jan Paul Medema^{2

3}, Giorgio Stassi¹

Affiliations

¹ Department of Surgical Oncological and Stomatological Sciences, University of Palermo, Palermo 90127, Italy.
² 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.
³ Oncode Institute, Amsterdam UMC, University of Amsterdam, Amsterdam 1105 AZ, the Netherlands.

Abstract

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).

Keywords: Cancer; Cell Biology; Cell isolation; Cell-based Assays; Flow Cytometry/Mass Cytometry; Molecular/Chemical Probes; Signal Transduction; Stem Cells.

Publication types

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

MeSH terms

Colorectal Neoplasms* / chemistry
Colorectal Neoplasms* / pathology
Flow Cytometry / methods*
Humans
Neoplastic Stem Cells* / chemistry
Neoplastic Stem Cells* / cytology
Neoplastic Stem Cells* / pathology
Wnt Signaling Pathway / genetics