Main description:

This volume provides detailed methods on the mechanisms of underlying cancer cell biology. Chapters guide readers through techniques for culturing cancer cell lines, xenografts, cryopreservation of tumor cells, analyzing the co-culture of breast cancer cells, protein secretion by ELISA, flow cytometry-based, multi-parametric immunofluorescence analysis, protein expression by western blot, analysis of surface protein levels, protein recycling by biotinylation assay, and proteomics analysis by liquid chromatography-mass spectrometry. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Cancer Cell Biology: Methods and Protocols aims to provide a

comprehensive set of tools for the analysis of cancer cell biology in the lab.

Contents:

1. Overview of the Maintenance of Authentic Cancer Cell Lines

Sherri L Christian

2. Culturing Suspension Cancer Cell Lines

Modeline N. Longjohn, Hong-Dien Phan, and Sherri L. Christian

3. Culture of Adherent Cancer Cell Lines

V. Noel and M.D. Berry

4. Breast Cancer Xenograft Murine Models

Wasundara Fernando, Krysta M. Coyle, and Paola Marcato

5. Processing and Cryopreservation of Blood, Cancer Tissues, and Cancer Cells For Viable Biobanking

Jennifer A. Chan and Suzanne M. Vercauteren

6. Adipocyte--Breast Cancer Cell Co--Culture In Transwells

Soofiyah P Ayaani, Martin Wabitsch, and Mark Christian

7. A 3-dimensional Co-Culture Method for Studying Interactions Between Adipocytes, Extracellular Matrix, and Cancer Cells

Emmanuel C. Asante, Nikitha K. Pallegar, Alicia M. Viloria-Petit, and Sherri L. Christian

8. Modelling Collective Invasion and Single-Cell Mesenchymal Invasion in Three-Dimensional Matrigel-Collagen I Cultures

Erin M. Bell, Marcia L. Graves, Pamela M. Dean, Tate O. Goodman, and Calvin D. Roskelley

9. CellTrace (TM) Violet Flow Cytometric Assay to Assess Cell Proliferation

Marina B. Lemieszek, Scott D. Findlay, and Gabrielle M. Siegers

10. Enzyme-Linked Immunosorbent Assay (ELISA)

Mahdis Sadat Tabatabaei and Marya Ahmed

11. Flow Cytometry Analysis to Detect Lapatinib-Induced Modulation of Constitutive and IFN-g induced HLA class I expression in HER2-positive Breast Cancer Cells

Louisa L. Wiede and Sheila Drover

12. An End to End Workflow for Interrogating Tumour Infiltrating Myeloid Cells Using Mass Cytometry

Maunish Barvalia and Kenneth W. Harder

13. Manual Immunofluorescence of Formalin-Fixed Paraffin-Embedded Human Tumour Tissues

Sarah Nersesian and Jeanette E. Boudreau

14. Western Blot Analysis of Lapatinib-Mediated Inhibition of the Epidermal Growth Factor Receptor 2 (HER2) Pathway in Breast Cancer Cells

Louisa L. Wiede and Sheila Drover

15. Evaluating Cell Membrane Localization and Intracellular Transport of Proteins by Biotinylation

Eduardo Reyes-Alvarez, Timothy J. Walker, and Lois M. Mulligan

16. Multiplexed Quantitative Proteomic Profiling of Cancer Cells and Tissues Using Isobaric Labeling-Based Tags

Mukhayyo Sultonova, Devanand M. Pinto, and J. Patrick Murphy

17. The use of Seahorse XF Assays to Interrogate Real-Time Energy Metabolism in Cancer Cell Lines

Jenna K. Caines, David A. Barnes, and Mark D. Berry

18. Using CRISPR-Cas9 to Dissect Cancer Mutations in Cell Lines

Shady Sayed, Duran Surun, Jovan Mircetic, Olga Sidorova, and Frank Buchholz

19. Single-cell RNA Sequencing Analysis using Fluidigm C1 platform for Characterization of Heterogeneous Transcriptomes

Jinhong Kim and Paola A. Marignani

20. Sequences to Differences in Gene Expression: Analysis of RNA-Seq Data

Polina V. Pavlovich and Pierre Cauchy

21. Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) for Gene Expression Analyses

Albert Caballero-Solares, Jennifer R. Hall, Xi Xue, and Matthew L. Rise

22. Peptide-affinity Isolation of Extracellular Vesicles and cell from Human Plasma -free DNA

Jeremy W. Roy, Catherine A. Taylor, Rodney J. Ouellette, and Stephen M. Lewis

23. Characterizing Extracellular vesicles using Nanoparticle Tracking Analysis

Modeline N. Longjohn and Sherri L. Christian