Skip to main content
SpringerLink
Log in

Hidden stressors in the clonogenic assay used in radiobiology experiments

  • Scientific Paper
  • Published:
Australasian Physical & Engineering Sciences in Medicine Aims and scope Submit manuscript

Abstract

While clonogenic assays are extensively used in radiobiology, there is no widely accepted procedure for choosing the composition of the cell culture media. Cell line suppliers recommend a specific culture medium for each cell line, however a researcher will frequently customize this aspect of the protocol by supplementing the recommended support medium with additives. For example, many researchers add antibiotics, in order to avoid contamination of cells and the consequent loss of data, with little discussion of the influence of the antibiotics on the clonogenic survival of the cells. It is assumed that the effect of any variables in the growth medium on cell survival is taken into consideration by comparing the survival fraction relative to that of controls grown under the same conditions. In the search for better cancer treatment, the effect of various stressors on clonogenic cell survival is under investigation. This study seeks to identify and test potential stressors commonly introduced into the cell culture medium, which may confound the response to radiation.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Elkind MM, Whitmore GF (1967) The radiobiology of cultured mammalian cells. Gordon and Breach, Paris

    Google Scholar 

  2. Hall EJ, Garcia AJ (2006) Radiobiology for the radiologist, Chap 3: Cell survival curves. Lippincott Williams & Wilkins, Philadelphia

    Google Scholar 

  3. Barber PR, Vojnovic B, Kelly J, Mayes CR, Boulton P, Woodcock M, Joiner MC (2001) Automated counting of mammalian cell colonies. Phys Med Biol 46:63–76

    Article  PubMed  CAS  Google Scholar 

  4. Bewes JM, Suchowerska N, McKenzie DR (2008) Automated cell colony counting and analysis using the circular Hough image transform algorithm (CHiTA). Phys Med Biol 53:5991–6008

    Article  PubMed  CAS  Google Scholar 

  5. Claridge Mackonis E, Suchowerska N, Zhang M, Ebert M, McKenzie DR, Jackson M (2007) Cellular response to modulated radiation fields. Phys Med Biol 52:5469–5482

    Article  PubMed  Google Scholar 

  6. Herok R, Konopacka M, Polanska J, Swierniak A, Rogolinski J, Jaksik R, Hancock R, Rzesowska-Wolny J (2010) Bystander effects induced by medium from irradiated cells: Similar transcriptome responses in irradiated and bystander K562 cells. Int J Radiat Oncol 77:244–252

    Article  CAS  Google Scholar 

  7. Lincoln K, Gabridge M (1998) Cell culture contamination: sources, consequences, prevention and elimination. Methods Cell Biol 57:49–65

    Article  PubMed  CAS  Google Scholar 

  8. McGarrity G (1976) Spread and control of mycoplasma infection of cell cultures. In Vitro 12:643–647

    Article  PubMed  CAS  Google Scholar 

  9. Zhou H, Suzuki M, Geard CR, Hei TK (2002) Effects of irradiated medium with or without cells on bystander cell responses. Mutat Res 499:135–141

    Article  PubMed  CAS  Google Scholar 

  10. Mothersill C, Seymour CB (1997) Medium from irradiated human epithelial cells but not human fibroblasts reduces the clonogenic survival of non-irradiated cells. Int J Radiat Biol 71:421–427

    Article  PubMed  CAS  Google Scholar 

  11. Suchowerska N, Ebert MA, Zhang M, Jackson M (2005) In vitro response of tumour cells to non-uniform irradiation. Phys Med Biol 50:3041–3051

    Article  PubMed  CAS  Google Scholar 

  12. Franken NAP, Rodermond HM, Stap J, Haveman J, van Bree C (2006) Clonigenic assay of cells in vitro. Nat Protoc 1:2315–2319

    Article  PubMed  CAS  Google Scholar 

  13. Claridge Mackonis E, Suchowerska N, Naseri P, McKenzie DR (2011) Optimisation of exposure conditions for in vitro (in press)

  14. International Atomic Energy Agency (2000) Absorbed dose determination in external beam radiotherapy. Technical report series no 398 IAEA Vienna, pp 210–214

  15. Hall EJ, Garcia AJ (2006) Radiobiology for the radiologist, Chap 28: Hyperthermia. 2006. Lippincott Williams & Wilkins, Philadelphia

    Google Scholar 

  16. Stostedt S, Bezak E (2010) Non-targeted effects of ionising radiation and radiotherapy. Australas Phys Eng Sci Med 33(3):219–231

    Article  Google Scholar 

Download references

Acknowledgment

We acknowledge the support of the NSW Cancer Council.

Author information

Authors and Affiliations

  1. Faculty of Medicine, University of Sydney, Sydney, NSW, Australia

    M. D. E. Potter

  2. Department of Radiation Oncology, Royal Prince Alfred Hospital, Missenden Road, Camperdown, Sydney, NSW, 2050, Australia

    M. D. E. Potter, N. Suchowerska & S. Rizvi

  3. School of Physics, University of Sydney, Sydney, NSW, Australia

    N. Suchowerska & D. R. McKenzie

Authors
  1. M. D. E. Potter
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Suchowerska
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. Rizvi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. R. McKenzie
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to N. Suchowerska.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Potter, M.D.E., Suchowerska, N., Rizvi, S. et al. Hidden stressors in the clonogenic assay used in radiobiology experiments. Australas Phys Eng Sci Med 34, 345–350 (2011). https://doi.org/10.1007/s13246-011-0082-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13246-011-0082-4

Keywords

Search

Navigation