Protocol for the derivation of primary cancer stem cell lines from human ependymal tumors

Cory M Richman; Peter B Dirks; Michael D Taylor; Kulandaimanuvel Antony Michealraj

doi:10.1016/j.xpro.2024.103260

Protocol for the derivation of primary cancer stem cell lines from human ependymal tumors

STAR Protoc. 2024 Sep 20;5(3):103260. doi: 10.1016/j.xpro.2024.103260. Epub 2024 Aug 15.

Authors

Cory M Richman¹, Peter B Dirks², Michael D Taylor³, Kulandaimanuvel Antony Michealraj⁴

Affiliations

¹ The Arthur and Sonia Labatt Brain Tumor Research Center, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Developmental & Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada. Electronic address: cory.richman@mail.utoronto.ca.
² The Arthur and Sonia Labatt Brain Tumor Research Center, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Developmental & Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Division of Neurosurgery, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada.
³ The Arthur and Sonia Labatt Brain Tumor Research Center, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Developmental & Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada; Department of Pediatrics - Hematology/Oncology, Baylor College of Medicine, Houston, TX 77030, USA; Cancer and Hematology Center, Texas Children's Hospital, Houston, TX 77030, USA; Dan L Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX 77030, USA.
⁴ The Arthur and Sonia Labatt Brain Tumor Research Center, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Developmental & Stem Cell Biology Program, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Neurological Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. Electronic address: micheala@pitt.edu.

Abstract

Cancer stem cells (CSCs) established from surgical biopsies closely mimic the human context and can be used to investigate disease mechanisms, genetic fitness, and therapeutic evaluation. Here, we present a protocol for the derivation of primary patient-derived CSC lines from ependymal tumors. We describe the necessary steps, from surgical intervention and biopsy to the dissociation of ependymomas to derive cultures. We then detail procedures for cell line propagation and define the characteristics of these primary cancer cell lines. For complete details on the use and execution of this protocol, please refer to Michealraj et al.¹.

Keywords: Cancer; Cell Biology; Cell Differentiation; Stem Cells.

MeSH terms

Brain Neoplasms / pathology
Cell Culture Techniques / methods
Cell Line, Tumor
Ependymoma / genetics
Ependymoma / pathology
Humans
Neoplastic Stem Cells* / metabolism
Neoplastic Stem Cells* / pathology