This detailed volume compiles the best methodologies and experimental techniques to profile and extract maximal data from brain tumor stem cells (BTSCs), the experimental paradigm for brain cancer research that offers insights into cancer stem cell populations that may drive not only tumor initiation but tumor recurrence and patient relapse. The BTSC model recapitulates scientific observations made in brain cancer patients, and these chapters provide the reader with a comprehensive understanding of the skills and techniques that will unlock data from this most informative subset of cells. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
1. Introduction to Brain Tumor Stem Cells
Nicolas Yelle, David Bahkshinyan, Chitra Venugopal, and Sheila K. Singh
2. Isolation and Culture of Glioblastoma Brain Tumor Stem Cells
Charles Chesnelong, Ian Restall, and Samuel Weiss
3. Establishment and Culture of Patient-Derived Primary Medulloblastoma Cell Lines
Sara Badodi, Silvia Marino, and Loredana Guglielmi
4. Bioinformatic Strategies for the Genomic and Epigenomic Characterization of Brain Tumors
Vijay Ramaswamy and Michael D. Taylor
5. Detecting Stem Cell Marker Expression Using the NanoString nCounter System
Scott Ryall, Anthony Arnoldo, Javal Sheth, Sheila K. Singh, and Cynthia Hawkins
6. Flow Cytometric Analysis of Brain Tumor Stem Cells
Minomi K. Subapanditha, Ashley A. Adile, Chitra Venugopal, and Sheila K. Singh
7. In Vitro Self-Renewal Assays for Brain Tumor Stem Cells
Mathieu Seyfrid, David Bobrowski, David Bakhshinyan, Nazanin Tatari, Chitra Venugopal, and Sheila K. Singh
8. Differentiation of Brain Tumor Initiating Cells
Michelle M. Kameda-Smith, Minomi K. Subapanditha, Sabra K. Salim, Chitra Venugopal, and Sheila K. Singh
9. The Study of Brain Tumor Stem Cell Migration
Montserrat Lara-Velázquez, Rawan Al-kharboosh, Luis Prieto, Paula Schiapparelli, and Alfredo Quiñones-Hinojosa
10. The Study of Brain Tumor Stem Cell Invasion
Rawan Al-kharboosh, Montserrat Lara-Velázquez, Luis Prieto, Rachel Sarabia-Estrada, and Alfredo Quiñones-Hinojosa
11. Cell Cycle Dynamics in Glioma Cancer Stem Cells
Ingrid Qemo and Lisa A. Porter
12. Embryonic Stem Cell Models of Human Brain Tumors
Ludivine Coudière Morrison, Nazanin Tatari, and Tamra Werbowetski-Ogilvie
13. Chromatin Immunoprecipitation (ChIP) Protocols for the Cancer and Developmental Biology Laboratory
Hunter McColl, Jamie L. Zagozewski, and David D. Eisenstat
14. EPH Profiling of BTIC Populations in Glioblastoma Multiforme Using CyTOF
Amy Hu, Jarrett J. Adams, Parvez Vora, Maleeha Qazi, Sheila Singh, Jason Moffat, and Sachdev S. Sidhu
15. Pooled Lentiviral CRISPR-Cas9 Screens for Functional Genomics in Mammalian Cells
Michael Aregger, Megha Chandrashekhar, Amy Tong, Kathrine Chan, and Jason Moffat
16. In Vitro Assays for Screening Small Molecules
Ashley A. Adile, David Bakhshinyan, Chitra Venugopal, and Sheila K. Singh
17. Drug Delivery in an Orthotopic Tumor Stem Cell-Based Model of Human Glioblastoma
Elena Binda, Alberto Visioli, Nadia Trivieri, and Angelo Luigi Vescovi
18. Engineering Inducible Knock-In Mice to Model Oncogenic Brain Tumor Mutations from Endogenous Loci
Jon D. Larson and Suzanne J. Baker
19. In Vivo Murine Models of Brain Metastasis
Mohini Singh, Neil Savage, and Sheila K. Singh
20. Cellular Magnetic Resonance Imaging for Tracking Metastatic Cancer Cells in the Brain
Katie M. Parkins, Ashley V. Makela, Amanda M. Hamilton, and Paula J. Foster
“Sheila K. Singh and Chitra Venugopal have developed a concise, systematic, and practical book; an ideal read for brain cancer researchers. … this book is recommended for anyone who strives for proficiency in biophysiology of cancer stem cells and hopes to discover novel therapeutic and treatment strategies for patients affected by devastating CNS disease.” (Angela M. Bohnen, Neurosurgery, Vol. 85 (4), October, 2019)
This detailed volume compiles the best methodologies and experimental techniques to profile and extract maximal data from brain tumor stem cells (BTSCs), the experimental paradigm for brain cancer research that offers insights into cancer stem cell populations that may drive not only tumor initiation but tumor recurrence and patient relapse. The BTSC model recapitulates scientific observations made in brain cancer patients, and these chapters provide the reader with a comprehensive understanding of the skills and techniques that will unlock data from this most informative subset of cells. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Includes cutting-edge techniques for the study of brain tumor stem cell research