Langdon Cancer Cell Culture

Methods and Protocols
Erscheinungsjahr 2008
ISBN: 978-1-59259-406-1
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 88, 360 Seiten, eBook

Reihe: Methods in Molecular Medicine

The culture of cancer cells is routinely practiced in many academic research centers, biotechnology companies, and hospital laboratories. Cancer Cell Culture: Methods and Protocols describes easy-to-follow methods to guide both novice and more experienced researchers seeking to use new techniques in their laboratories. Our present understanding of the cell and molecular biology of cancer has been derived mainly from the use of cultured cancer cells and we cover a number of the most widely used assays to study function in current use. Part I introduces the basic concept of cancer cell culture and this is followed by a description of the general techniques used in many cell culture facilities. The importance of cell line characterization is now widely recognized and methods to characterize and authenticate cell lines are described in Part II. Part III covers the isolation and development of specific cancer cell types and provides valuable tips for those wishing to derive new cell line models. A wide range of procedures encompassing many of the key functional features of cancer cells are described in Part IV including assays to evaluate clonogenicity, cell proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and cell cycle parameters. Methods to modify cancer cells are described in Part V, including protocols for transfection, development of drug-resistance, immortalization, and transfer in vivo. In Part VI methods of coculture of different cell types and contamination of cell lines are covered.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Langdon, Simon P.

Inhaltsverzeichnis

to Cancer Cell Culture.- Basic Principles of Cancer Cell Culture.- Essential Techniques of Cancer Cell Culture.- Characterization and Authentication.- Characterization and Authentication of Cancer Cell Lines.- Authentication of Cancer Cell Lines by DNA Fingerprinting.- Cytogenetic Characterization of Tumor Cell Lines.- Isolation and Culture of Specialized Cell Types.- Isolation and Culture of Colon Cancer Cell Lines.- Isolation and Culture of Melanoma Cell Lines.- Isolation and Culture of Human Brain Tumor Cells.- Isolation and Culture of Renal Cancer Cell Lines.- Isolation and Culture of Prostate Cancer Cell Lines.- Isolation and Culture of Ovarian Cancer Cell Lines.- Isolation and Culture of Leukemia Cell Lines.- Functional Assays in Cell Culture.- Cell Sensitivity Assays: Clonogenic Assay.- Cell Sensitivity Assays: The MTT Assay.- PARP Cleavage as a Means of Assessing Apoptosis.- Detection of Apoptosis by the TUNEL Assay.- Apoptosis Measurement by Annexin V Staining.- Cell Adhesion Assays.- Radial Monolayer Cell Migration Assay.- Invasion and Motility Assays.- Identification of Senescence in Cancer Cells.- Angiogenesis Assays.- Flow Cytometric DNA Analysis of Human Cancer Cell Lines.- Modification of Cancer Cells.- DNA-Mediated Gene Transfer.- Development of Drug-Resistant Models.- Immortalization of Human Prostate Cells With the Human Papillomavirus Type 16 E6 Gene.- The Use of Matrigel to Facilitate the Establishment of Human Cancer Cell Lines as Xenografts.- Coculture Systems.- Coculture of Prostate Cancer Cells.- Coculture of Ovarian Cells Using Porous Tissue Culture Inserts.- Cell Line Contamination.- Cell Culture Contamination.- Detecting Mycoplasma Contamination in Cell Cultures by Polymerase Chain Reaction.- Elimination of Mycoplasma from Infected Cell LinesUsing Antibiotics.