Cancer Cell Culture

This new edition covers the recent advances in cancer cell culture research, including novel cell culture procedures. It includes methods of authenticating cell lines, functional assays to evaluate clonogenicity, and stresses safe working practice throughout.

With many recent advances, cancer cell culture research is more important than ever before. This timely edition of Cancer Cell Culture: Methods and Protocols covers the basic concepts of cancer cell biology and culture while expanding upon the recent shift in cell culture methods from the generation of new cell lines to the use of primary cells. There are methods to characterize and authenticate cell lines, to isolate and develop specific types of cancer cells, and to develop new cell line models. Functional assays are provided for the evaluation of clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters. Other methods permit the modification of cells for transfection, drug resistance, immortalization, and transfer in vivo, the co-culture of different cell types, and the detection and treatment of contamination. In this new edition, specific emphasis is placed on safe working practice for both cells and laboratory researchers. These chapters contain the information critical to success only by good practice and quality control will the results of cancer cell culture improve. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Cancer Cell Culture: Methods and Protocols serves as a practical guide for scientists of all backgrounds and aims to convey the appropriate sense of fascination associated with this research field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Cancer Biology.- Principles of Cancer Cell Culture.- Storage of Cell Lines.- Characterization and Authentication of Cancer Cell Lines: An Overview.- Online Verification of Human Cell Line Identity by STR DNA Typing.- Cytogenetic Analysis of Cancer Cell Lines.- Cell Culture Contamination.- Detecting Mycoplasma Contamination in Cell Cultures by Polymerase Chain Reaction.- Elimination of Mycoplasma from Infected Cell Lines Using Antibiotics.- Quality Assurance and Good Laboratory Practice.- Generation of Lung Cancer Cell Line Variants by Drug Selection or Cloning.- Isolation and Culture of Colon Cancer Cells and Cell Lines.- Isolation and Culture of Melanoma and Naevus Cells and Cell Lines.- Isolation and Culture of Squamous Cell Carcinomalines.- Isolation and Culture of Ovarian Cancer Cells and Cell Lines.- Establishment and Culture of Leukemia-Lymphoma Cell Lines.- Isolation of Inflammatory Cells from Human Tumors.- Isolation of Endothelial Cells from Human Tumors.- Cellular Chemosensitivity Assays: An Overview.- Cell Sensitivity Assays: The MTT Assay.- Cell Sensitivity Assays: The ATP-based Tumor Chemosensitivity Assay.- The Differential Staining Cytotoxicity (DiSC) Assay: A Review.- Real-time Cytotoxicity Assays.- FPurification of Annexin V and its use in the Detection of Apoptotic Cells.- Measurement of DNA Damage in Individual Cells Using the Single Cell Gel Electrophoresis (Comet) Assay.- Molecular Breakpoint Analysis of Chromosome Translocations in Cancer Cell Lines by Long Distance Inverse-PCR.- Cell Migration and Invasion Assays.- Angiogenesis Assays.- Flow Cytometric DNA Analysis of Human Cancers and Cell Lines.- Expression Analysis of Homeobox Genes in Leukemia/Lymphoma Cell Lines.- Measuring Gene Expression from Cell Cultures by Quantitative Reverse-Transciptase Polymerase Chain Reaction.- Proteomic Evaluation of Cancer Cells Identification of Cell Surface Proteins.- Development of Rituximab-resistant B-NHL Clones:An In Vitro Model for Studying Tumor Resistance to Monoclonal Antibody-mediated Immunotherapy.- High Throughput Screening of Natural Products for Cancer Therapy: A Review.- Analysis of Drug Interactions.- Transfection and DNA-mediated Gene Transfer.- Drug Design and Testing: Profiling of Anti-proliferative Agents for Cancer Therapy Using a Cell-based Methyl-[3H]-Thymidine Incorporation Assay.- Feeder Layers: Co-culture with Non-neoplastic Cells.- Xenotransplantation of Breast Cancers