Cancer Cell Culture: Methods and Protocols / Edition 1

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Overview

Expert researchers describe in detail their most productive and up-to-date methods for growing cancer cells in the laboratory. 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 to evaluate clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters. Other methods permit the modification of cancer cells for transfection, development of drug resistance, immortalization, and transfer in vivo; the co-culture of different cell types; and the detection and treatment of contamination.

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Editorial Reviews

Doody's Review Service
Reviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This is a compilation of protocols for the isolation, characterization, and culture of tumor cells. Additional material includes identification of contaminants, composition of media, and sources of cell lines.
Purpose: The goal of this book is to provide investigators with well established methods for culturing tumor cells. General principles as well as specific procedures are described. This is a generally useful objective, but there are limitations in presentation.
Audience: This is basically a laboratory manual and as such is intended for advanced students, research fellows, and experienced laboratory staff. The multiple authors have documented expertise in the area.
Features: The ability to culture transformed cells in the laboratory has permitted many detailed studies of cell behavior, gene expression, growth regulation, etc., that would otherwise be difficult, if not impossible. Hence, a compilation of protocols focused on this area has obvious currency. This book is divided into seven sections and has 32 chapters plus two appendixes. The initial sections provide some general techniques and include material on characterization. This is followed by a series of chapters on isolation and line establishment of cells from several tumor types and a set of assays generally applied to such cell lines. Protocols for transfection and co-culture are included, and the book concludes with several chapters on contamination. In general, protocols provide sufficient detail for adaptation in the laboratory. In several cases, however, the material would have benefited from inclusion of a discussion on the shortcomings of a given assay, limitations, and reproducibility (angiogenesis assays, for example). In addition, the genetic stability of cultured tumor cells is limited, especially with continued passage in the laboratory. In this arena, the importance of genetic characterization received insufficient emphasis. It is well recognized that all such cultured systems have limitations — here, a candid discussion of these juxtaposed with the use of cell lines for toxicity and drug screening would have been useful. Finally, the appendixes provide a listing of ingredients of commonly used media (not too useful since virtually all investigators buy prepared medium) and a listing of available cell lines from central repositories (easily obtained from the respective Web sites). Laboratories wishing to have a handy cell culture protocol set will find value in this contribution.
Assessment: This is a book of mixed quality. Stronger editing and the inclusion of more critical evaluations of methods would have helped. The approaches are somewhat dated. Many of the procedures are standard and used in diverse areas; little unique material is presented.
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Product Details

  • ISBN-13: 9781588290793
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 1/5/2010
  • Series: Methods in Molecular Medicine Series , #88
  • Edition number: 1
  • Pages: 368
  • Product dimensions: 9.21 (w) x 6.14 (h) x 0.88 (d)

Table of Contents

Part I. Introduction to Cancer Cell Culture

Basic Principles of Cancer Cell Culture Simon P. Langdon

Essential Techniques of Cancer Cell Culture Kenneth G. Macleod and Simon P. Langdon

Part II. Characterization and Authentication

Characterization and Authentication of Cancer Cell Lines: An Overview Simon P. Langdon

Authentication of Cancer Cell Lines by DNA Fingerprinting Wilhelm G. Dirks and Hans G. Drexler

Cytogenetic Characterization of Tumor Cell Lines Roderick A. F. MacLeod and Hans G. Drexler

Part III. Isolation and Culture of Specialized Cell Types

Isolation and Culture of Colon Cancer Cell Lines Jae-Gahb Park, Ja-Lok Ku, and So-Yeon Park

Isolation and Culture of Melanoma Cell Lines Adam I. Riker

Isolation and Culture of Human Brain Tumor Cells German G. Gomez and Carol A. Kruse

Isolation and Culture of Renal Cancer Cell Lines Jae-Gahb Park, Ja-Lok Ku, and So-Yeon Park

Isolation and Culture of Prostate Cancer Cell Lines Nora M. Navone, Matilde Olive, and Patricia Troncoso

Isolation and Culture of Ovarian Cancer Cell Lines Simon P. Langdon

Isolation and Culture of Leukemia Cell Lines Hans G. Drexler

Part IV. Functional Assays in Cell Culture

Cell Sensitivity Assays: Clonogenic Assay Jane A. Plumb

Cell Sensitivity Assays: The MTT Assay Jane A. Plumb

PARP Cleavage as a Means of Assessing Apoptosis Peter Mullen

Detection of Apoptosis by the TUNEL Assay John Lawry

Apoptosis Measurement by Annexin V Staining Eric Miller

Cell Adhesion Assays William N. Scott

Radial Monolayer Cell Migration Assay Michael E. Berens and Christian Beaudry

Invasion and Motility Assays William N. Scott

Identification of Senescence in Cancer Cells Lesley J. Jardine

Angiogenesis Assays William N. Scott

Flow Cytometric DNA Analysis of Human Cancer Cell Lines Peter Mullen

Part V. Modification of Cancer Cells

DNA-Mediated Gene Transfer Amanda J. M. O'Donnell

Development of Drug-Resistant Models Helen M. Coley

Immortalization of Human Prostate Cells With the Human Papillomavirus Type 16 E6 Gene Norman J. Maitland, Catherine A. Macintosh, Christian Schmitz, and Shona H. Lang

The Use of Matrigel to Facilitate the Establishment of Human Cancer Cell Lines as Xenografts Peter Mullen

Part VI. Coculture Systems

Coculture of Prostate Cancer Cells Shona H. Lang and Norman J. Maitland

Coculture of Ovarian Cells Using Porous Tissue Culture Inserts Katherine E. Snow and Simon P. Langdon

Part VII. Cell Line Contamination

Cell Culture Contamination: An Overview Simon P. Langdon

Detecting Mycoplasma Contamination in Cell Cultures by Polymerase Chain Reaction Cord C. Uphoff and Hans G. Drexler

Elimination of Mycoplasma from Infected Cell Lines Using Antibiotics Cord C. Uphoff and Hans G. Drexler

Appendix 1: Formulations of Commonly Used Cell Culture Media

Appendix 2: Human Cancer Cell Lines Available from the ATCC and DSMZ Cell Banks (Combined List)

Index

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