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

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.

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Affiliated Institutions

• Ann Arbor Center for Independent Living US
• University of Gothenburg SE
• University of Graz AT
• Universität Ulm DE
• University of Massachusetts Chan Medical School US
• Trinity College Dublin IE
• Royal College of Surgeons in Ireland IE
• Yale University US
• University of Padua IT
• Aix-Marseille Université FR
• University College London GB
• University of Göttingen DE
• National University of Singapore SG
• Merck & Co., Inc., Rahway, NJ, USA (United States) US
• Technion – Israel Institute of Technology IL
• Centre de Gestion Scientifique FR
• La Trobe University AU
• University of Pennsylvania US
• Howard Hughes Medical Institute US
• Duke University US
• Icahn School of Medicine at Mount Sinai US
• University of North Carolina at Chapel Hill US
• Harvard University Press US
• Université Claude Bernard Lyon 1 FR
• Roswell Park Comprehensive Cancer Center US
• University of Amsterdam NL
• Karolinska Institutet SE
• Istituto Superiore di Sanità IT
• University of Rome Tor Vergata IT
• Kyoto University JP
• University of Leicester GB
• Philadelphia University US
• Austrian Academy of Sciences AT
• Amsterdam University of the Arts NL
• Universidad de Buenos Aires AR
• University of Freiburg DE
• Université de Bourgogne FR
• The University of Texas Southwestern Medical Center US
• Université de Strasbourg FR
• University of Würzburg DE
• Boston Children's Hospital US
• University of Zurich CH
• University of Cambridge GB
• Louisiana State University Health Sciences Center New Orleans US
• Université Paris-Sud FR
• University of Geneva CH
• Rockefeller University US
• University of Chieti-Pescara IT
• University of Lisbon PT
• Ghent University Hospital BE
• Weizmann Institute of Science IL
• National Institutes of Health US
• Thomas Jefferson University US
• University of Glasgow GB
• University of Chicago US
• Inserm FR
• Boston Children's Museum US
• University of California, San Diego US
• University of Michigan US
• University of Bern CH
• New York University US
• Institut Pasteur FR
• McMaster University CA
• Buck Institute for Research on Aging US
• Johns Hopkins University US
• University of California, San Francisco US
• The University of Tokyo JP
• Stony Brook University US
• The University of Osaka JP
• University of Lausanne CH

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