Cancer Cell Biology

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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt
Overview of the maintenance of authentic cancer cell lines.- Culturing suspension cancer cell lines.- Culture of Adherent Cancer Cell Lines.- Breast Cancer Xenograft Murine Models.- Processing and cryopreservation of blood, cancer tissues and cancer cells for viable biobanking.- Adipocyte-breast cancer cell co-culture in transwells.- A 3-dimensional co-culture method for studying interactions between adipocytes, extracellular matrix, and cancer cells.- Modelling Collective Invasion and Single-Cell Mesenchymal Invasion in Three-Dimensional Matrigel-Collagen I Cultures.- CellTrace™ Violet Flow Cytometric Assay to Assess Cell Proliferation.- Enzyme-Linked Immunosorbent Assay (ELISA).- Flow cytometry analysis to detect lapatinib-induced modulation of constitutive and IFN-g induced HLA class I expression in HER2-positive breast cancer cells.- An end-to-end workflow for interrogating tumour infiltrating myeloid cells using mass cytometry.- Manual Immunofluorescence of Formalin-Fixed Paraffin-Embedded Human Tumour Tissues.- Western blot analysis of lapatinib-mediated inhibition of the epidermal growth factor receptor 2 (HER2) pathway in breast cancer cells.- Evaluating cell membrane localization and intracellular transport of proteins by biotinylation.- Multiplexed quantitative proteomic profiling of cancer cells and tissues using isobaric labeling-based tags.- The use of Seahorse XF assays to interrogate real-time energy metabolism in cancer cell lines.- Using CRISPR-Cas9 to dissect cancer mutations in cell Iines.- Single-cell RNA sequencing analysis using Fluidigm C1 platform for characterization of heterogeneous transcriptomes.- Sequences to Differences in Gene Expression: Analysis of RNA-Seq Data.- Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) for gene expression analyses.- Peptide-affinity isolation of extracellular vesicles and cell-free DNA from human plasma.- Characterizing Extracellularvesicles using Nanoparticle Tracking Analysis.