Cancer Cell Culture: Methods and Protocols

This volume describes new techniques for the culture of cancer cells. The first section introduces the rationale behind selection of specific materials to help the researcher select culture conditions appropriate to their studies and discusses general techniques operating in many culture facilities. The second section covers specific requirements of individual cancer cell types for optimal growth and maintenance. A range of procedures are described in section three which include assays to evaluate proliferation, viability, cytotoxicity, apoptosis, migration, invasion, and angiogenesis. Techniques of gene transfer and the development of drug resistance are also described. The book concludes with methods of co-culture of different cell types.

PART I . INTRODUCTION TO CANCER CELL CULTURE
1. Basic principles of cancer cell culture
2. Essential techniques of cancer cell culture
PART II. CHARACTERIZATION AND AUTHENTICATION
3. Characterization and authentication of cancer cell lines: An overview
4. Authentication of cancer cell lines by DNA fingerprinting
5. Cytogenetic characterization of tumor cell lines
PART III. ISOLATION AND CULTURE OF SPECIALIZED CELL TYPES
6. Isolation and culture of colon cancer cell lines
7. Isolation and culture of melanoma cell lines
8. Isolation and culture of human brain tumour cells
9. Isolation and culture of renal cancer cell lines
10. Isolation and culture of prostate cancer cell lines
11. Isolation and culture of ovarian cancer cell lines
12. Isolation and culture of leukemia cell lines
PART IV. FUNCTIONAL ASSAYS IN CELL CULTURE
13 Cell sensitivity assays: Clonogenic assay
14. Cell sensitivity assays : The MTT assay
15. PARP cleavage as a means of assessing apoptosis
16. Detection of apoptosis by the TUNEL assay
17. Apoptosis measurement by Annexin V staining
18. Cell adhesion assays
19. Radial Monolayer Cell Migration Assay
20. Invasion and motility assays
21. Identification of senescence in cancer cells
22. Angiogenesis assays
23. Flow cytometric DNA analysis of human cancer cell lines
PART V MODIFICATION OF CANCER CELLS
24. DNA-mediated gene transfer
25. Development of drug-resistant models
26. Immortalisation of human prostate cells with the human papillomavirus type 16E6 gene
27. The Use of Matrigel to Facilitate the Establishment of Human Carcinoma Cell Lines as Xenografts
PART VI CO-CULTURE SYSTEMS
28. Co-culture of prostate cancer cells
29. Co-culture of ovarian cancer cells

PART VII. CELL LINE CONTAMINATION
30. Cell culture contamination – an overview
31. Detecting Mycoplasma contamination in cell cultures by polymerase chain reaction
32. Elimination of Mycoplasma from infected cell lines using antibiotics
PART VIII. APPENDICES
Appendix 1. Formulations of commonly used cell culture media.
Appendix 2. Human cancer cell lines available from the ATCC and DSMZ
Index