Skip to main content

Cell Sensitivity Assays: The ATP-based Tumor Chemosensitivity Assay

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 731))

Abstract

The ATP-based tumor chemosensitivity assay (ATP–TCA) is a standardised system which can be adapted to a variety of uses with both cell lines and primary cell cultures. It has a strong track record in drug development, mechanistic studies of chemoresistance and as an aid to clinical decision-making. The method starts with the extraction of cells in suspension from continuous cell culture (for cell lines), malignant effusions or biopsy material. Enzymatic digestion is used to obtain cells from tumor tissue. The assay uses a serum-free medium and polypropylene plates to prevent the growth of non-neoplastic cells over a 6-day incubation period followed by detergent-based extraction of cellular ATP for measurement by luciferin–luciferase assay in a luminometer. The assay results are usually shown as percentage inhibition at each concentration tested, and can be used with suitable software to examine synergy between different anticancer agents.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Hunter, E. M., Sutherland, L. A., Cree, I. A., Dewar, J. A., Preece, P. E., Wood, R. A., Linder, D., and Andreotti, P. E. (1993) Heterogeneity of chemosensitivity in human breast carcinoma: use of an adenosine triphosphate (ATP) chemiluminescence assay, Eur J Surg Oncol 19, 242–249.

    PubMed  CAS  Google Scholar 

  2. Andreotti, P. E., Cree, I. A., Kurbacher, C. M., Hartmann, D. M., Linder, D., Harel, G., Gleiberman, I., Caruso, P. A., Ricks, S. H., Untch, M., Sartori, D.C. and Bruckner H. (1995) Chemosensitivity testing of human tumors using a microplate adenosine triphosphate luminescence assay: clinical correlation for cisplatin resistance of ovarian carcinoma, Cancer Res 55, 5276–5282.

    PubMed  CAS  Google Scholar 

  3. Cree, I. A., Kurbacher, C. M., Untch, M., Sutherland, L. A., Hunter, E. M., Subedi, A. M., James, E. A., Dewar, J. A., Preece, P. E., Andreotti, P. E., and Bruckner, H. W. (1996) Correlation of the clinical response to chemotherapy in breast cancer with ex vivo chemosensitivity, Anticancer Drugs 7, 630–635.

    Article  PubMed  CAS  Google Scholar 

  4. Sharma, S., Neale, M. H., Di Nicolantonio, F., Knight, L. A., Whitehouse, P. A., Mercer, S. J., Higgins, B. R., Lamont, A., Osborne, R., Hindley, A. C., Kurbacher, C. M., and Cree, I. A. (2003) Outcome of ATP-based tumor chemosensitivity assay directed chemotherapy in heavily pre-treated recurrent ovarian carcinoma, BMC Cancer 3, 19.

    Article  PubMed  Google Scholar 

  5. Cree, I. A., Kurbacher, C. M., Lamont, A., Hindley, A. C., and Love, S. (2007) A prospective randomized controlled trial of tumour chemosensitivity assay directed chemotherapy versus physician’s choice in patients with recurrent platinum-resistant ovarian cancer, Anticancer Drugs 18, 1093–1101.

    Article  PubMed  CAS  Google Scholar 

  6. Konecny, G., Crohns, C., Pegram, M., Felber, M., Lude, S., Kurbacher, C., Cree, I. A., Hepp, H., and Untch, M. (2000) Correlation of drug response with the ATP tumorchemosensitivity assay in primary FIGO stage III ovarian cancer, Gynecol Oncol 77, 258–263.

    Article  PubMed  CAS  Google Scholar 

  7. Ugurel, S., Schadendorf, D., Pfohler, C., Neuber, K., Thoelke, A., Ulrich, J., Hauschild, A., Spieth, K., Kaatz, M., Rittgen, W., Delorme, S., Tilgen, W., and Reinhold, U. (2006) In vitro drug sensitivity predicts response and survival after individualized sensitivity-directed chemotherapy in metastatic melanoma: a multicenter phase II trial of the Dermatologic Cooperative Oncology Group, Clin Cancer Res 12, 5454–5463.

    Article  PubMed  CAS  Google Scholar 

  8. Kurbacher, C. M., and Cree, I. A. (2005) Chemosensitivity testing using microplate adenosine triphosphate-based luminescence measurements, Methods Mol Med 110, 101–120.

    PubMed  CAS  Google Scholar 

  9. Kurbacher, C. M., Bruckner, H. W., Cree, I. A., Kurbacher, J. A., Wilhelm, L., Poch, G., Indefrei, D., Mallmann, P., and Andreotti, P. E. (1997) Mitoxantrone combined with paclitaxel as salvage therapy for platinum-refractory ovarian cancer: laboratory study and clinical pilot trial, Clin Cancer Res 3, 1527–1533.

    PubMed  CAS  Google Scholar 

  10. Myatt, N., Cree, I. A., Kurbacher, C. M., Foss, A. J., Hungerford, J. L., and Plowman, P. N. (1997) The ex vivo chemosensitivity profile of choroidal melanoma, Anticancer Drugs 8, 756–762.

    Article  PubMed  CAS  Google Scholar 

  11. Neale, M. H., Myatt, N., Cree, I. A., Kurbacher, C. M., Foss, A. J., Hungerford, J. L., and Plowman, P. N. (1999) Combination chemotherapy for choroidal melanoma: ex vivo sensitivity to treosulfan with gemcitabine or cytosine arabinoside, Br J Cancer 79, 1487–1493.

    Article  PubMed  CAS  Google Scholar 

  12. Corrie, P. G., Shaw, J., Spanswick, V. J., Sehmbi, R., Jonson, A., Mayer, A., Bulusu, R., Hartley, J. A., and Cree, I. A. (2005) Phase I trial combining gemcitabine and treosulfan in advanced cutaneous and uveal melanoma patients, Br J Cancer 92, 1997–2003.

    Article  PubMed  CAS  Google Scholar 

  13. Pfohler, C., Cree, I. A., Ugurel, S., Kuwert, C., Haass, N., Neuber, K., Hengge, U., Corrie, P. G., Zutt, M., Tilgen, W., and Reinhold, U. (2003) Treosulfan and gemcitabine in metastatic uveal melanoma patients: results of a multicenter feasibility study, Anticancer Drugs 14, 337–340.

    Article  PubMed  Google Scholar 

  14. Di Nicolantonio, F., Neale, M. H., Knight, L. A., Lamont, A., Skailes, G. E., Osborne, R. J., Allerton, R., Kurbacher, C. M., and Cree, I. A. (2002) Use of an ATP-based chemosensitivity assay to design new combinations of high-concentration doxorubicin with other drugs for recurrent ovarian cancer, Anticancer Drugs 13, 625–630.

    Article  PubMed  Google Scholar 

  15. Knight, L. A., Di Nicolantonio, F., Whitehouse, P., Mercer, S., Sharma, S., Glaysher, S., Johnson, P., and Cree, I. A. (2004) The in vitro effect of gefitinib (‘Iressa’) alone and in combination with cytotoxic chemotherapy on human solid tumours, BMC Cancer 4, 83.

    Article  PubMed  Google Scholar 

  16. Knight, L. A., Di Nicolantonio, F., Whitehouse, P. A., Mercer, S. J., Sharma, S., Glaysher, S., Hungerford, J. L., Hurren, J., Lamont, A., and Cree, I. A. (2006) The effect of imatinib mesylate (Glivec) on human tumor-derived cells, Anticancer Drugs 17, 649–655.

    Article  PubMed  CAS  Google Scholar 

  17. Di Nicolantonio, F., Knight, L. A., Glaysher, S., Whitehouse, P. A., Mercer, S. J., Sharma, S., Mills, L., Prin, A., Johnson, P., Charlton, P. A., Norris, D., and Cree, I. A. (2004) Ex vivo reversal of chemoresistance by tariquidar (XR9576), Anticancer Drugs 15, 861–869.

    Article  PubMed  Google Scholar 

  18. Knight, L. A., Kurbacher, C. M., Glaysher, S., Fernando, A., Reichelt, R., Dexel, S., Reinhold, U., and Cree, I. A. (2009) Activity of mevalonate pathway inhibitors against breast and ovarian cancers in the ATP-based tumour chemosensitivity assay, BMC Cancer 9, 38.

    Article  PubMed  Google Scholar 

  19. Fernando, A., Glaysher, S., Conroy, M., Pekalski, M., Smith, J., Knight, L. A., Di Nicolantonio, F., and Cree, I. A. (2006) Effect of culture conditions on the chemosensitivity of ovarian cancer cell lines, Anticancer Drugs 17, 913–919.

    Article  PubMed  CAS  Google Scholar 

  20. Di Nicolantonio, F., Mercer, S. J., Knight, L. A., Gabriel, F. G., Whitehouse, P. A., Sharma, S., Fernando, A., Glaysher, S., Di Palma, S., Johnson, P., Somers, S. S., Toh, S., Higgins, B., Lamont, A., Gulliford, T., Hurren, J., Yiangou, C., and Cree, I. A. (2005) Cancer cell adaptation to chemotherapy, BMC Cancer 5, 78.

    Article  PubMed  Google Scholar 

  21. Mercer, S. J., Di Nicolantonio, F., Knight, L. A., Gabriel, F. G., Whitehouse, P. A., Sharma, S., Fernando, A., Bhandari, P., Somers, S. S., Toh, S. K., and Cree, I. A. (2005) Rapid up-regulation of cyclooxygenase-2 by 5-fluorouracil in human solid tumors, Anticancer Drugs 16, 495–500.

    Article  PubMed  CAS  Google Scholar 

  22. Glaysher, S., Yiannakis, D., Gabriel, F. G., Johnson, P., Polak, M. E., Knight, L. A., Goldthorpe, Z., Peregrin, K., Gyi, M., Modi, P., Rahamim, J., Smith, M. E., Amer, K., Addis, B., Poole, M., Narayanan, A., Gulliford, T. J., Andreotti, P. E., and Cree, I. A. (2009) Resistance gene expression determines the in vitro chemosensitivity of non-small cell lung cancer (NSCLC), BMC Cancer 9, 300.

    Article  PubMed  Google Scholar 

  23. Hunter, E. M., Sutherland, L. A., Cree, I. A., Subedi, A. M., Hartmann, D., Linder, D., and Andreotti, P. E. (1994) The influence of storage on cytotoxic drug activity in an ATP-based chemosensitivity assay, Anti-cancer drugs 5, 171–176.

    Article  PubMed  CAS  Google Scholar 

  24. Palmgren, J. J., Monkkonen, J., Korjamo, T., Hassinen, A., and Auriola, S. (2006) Drug adsorption to plastic containers and retention of drugs in cultured cells under in vitro conditions, Eur J Pharm Biopharm 64, 369–378.

    Article  PubMed  CAS  Google Scholar 

  25. Di, S., Li-Fen, H., and Jessie, L. S. A. (1996) Binding of taxol to plastic and glass containers and protein under in vitro conditions, Journal of Pharmaceutical Sciences 85, 29–31.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ian A. Cree .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Glaysher, S., Cree, I.A. (2011). Cell Sensitivity Assays: The ATP-based Tumor Chemosensitivity Assay. In: Cree, I. (eds) Cancer Cell Culture. Methods in Molecular Biology, vol 731. Humana Press. https://doi.org/10.1007/978-1-61779-080-5_21

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-080-5_21

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-079-9

  • Online ISBN: 978-1-61779-080-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics