Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes

L. Galluzzi, S. A. Aaronson, J. Abrams, E. S. Alnemri, D. W. Andrews, E. H. Baehrecke, N. G. Bazan, M. V. Blagosklonny, K. Blomgren, C. Borner, D. E. Bredesen, C. Brenner, M. Castedo, J. A. Cidlowski, A. Ciechanover, G. M. Cohen, V. De Laurenzi, R. De Maria, M. Deshmukh, B. D. DynlachtW. S. El-Deiry, R. A. Flavell, S. Fulda, C. Garrido, P. Golstein, M. L. Gougeon, D. R. Green, H. Gronemeyer, G. Hajnóczky, J. M. Hardwick, M. O. Hengartner, H. Ichijo, M. Jäättelä, O. Kepp, A. Kimchi, D. J. Klionsky, R. A. Knight, S. Kornbluth, S. Kumar, B. Levine, S. A. Lipton, E. Lugli, F. Madeo, W. Malorni, J. C.W. Marine, S. J. Martin, J. P. Medema, P. Mehlen, G. Melino, U. M. Moll, E. Morselli, S. Nagata, D. W. Nicholson, P. Nicotera, G. Nuñez, M. Oren, J. Penninger, S. Pervaiz, M. E. Peter, M. Piacentini, J. H.M. Prehn, H. Puthalakath, G. A. Rabinovich, R. Rizzuto, C. M.P. Rodrigues, D. C. Rubinsztein, T. Rudel, L. Scorrano, H. U. Simon, H. Steller, J. Tschopp, Y. Tsujimoto, P. Vandenabeele, I. Vitale, K. H. Vousden, R. J. Youle, J. Yuan, B. Zhivotovsky, G. Kroemer

Research output: Contribution to journalReview articlepeer-review

578 Scopus citations

Abstract

Cell death is essential for a plethora of physiological processes, and its deregulation characterizes numerous human diseases. Thus, the in-depth investigation of cell death and its mechanisms constitutes a formidable challenge for fundamental and applied biomedical research, and has tremendous implications for the development of novel therapeutic strategies. It is, therefore, of utmost importance to standardize the experimental procedures that identify dying and dead cells in cell cultures and/or in tissues, from model organisms and/or humans, in healthy and/or pathological scenarios. Thus far, dozens of methods have been proposed to quantify cell death-related parameters. However, no guidelines exist regarding their use and interpretation, and nobody has thoroughly annotated the experimental settings for which each of these techniques is most appropriate. Here, we provide a nonexhaustive comparison of methods to detect cell death with apoptotic or nonapoptotic morphologies, their advantages and pitfalls. These guidelines are intended for investigators who study cell death, as well as for reviewers who need to constructively critique scientific reports that deal with cellular demise. Given the difficulties in determining the exact number of cells that have passed the point-of-no-return of the signaling cascades leading to cell death, we emphasize the importance of performing multiple, methodologically unrelated assays to quantify dying and dead cells.

Original languageEnglish
Pages (from-to)1093-1107
Number of pages15
JournalCell Death and Differentiation
Volume16
Issue number8
DOIs
StatePublished - 2009

Keywords

  • Apoptosis
  • Cell Death
  • Eukaryotic Cells/cytology
  • Flow Cytometry
  • Guidelines as Topic
  • Humans
  • Immunoblotting
  • Microscopy, Electron, Scanning
  • Microscopy, Fluorescence
  • Spectrometry, Fluorescence

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