Abstract
Cells can die via two distinct pathways, apoptosis and necrosis. Cells that are exposed to severe traumatizing conditions usually undergo an uncontrolled swelling and lyse via necrosis (1). However, a cell dying under less traumatizing conditions has limited control over its own demise and utilize a mode of programmed cell death (apoptosis) first characterized by Kerr et al. (2). It has been demonstrated that cells undergoing apoptosis usually share a number of common features, such as cell shrinkage, externalization of phosphatidylserine, nuclear condensation, nuclear fragmentation, and degradation of the DNA into nucleosomal fragments (3–5).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Trump, B. F. and Ginn F. L., (1969) The pathogenesis of subcellular reaction to lethal injury, in Methods and Achievements in Experimental Pathology, vol. 4 (Bajusz, E. and Jasmin, G., eds.), Karger, Basel, pp. 1–10.
Kerr, J. F. R., Wyllie, A. H., and Currie, A. R. (1972) Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics. Br. J. Cancer 26, 239–257.
Wyllie, A. H. (1980) Glutocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature Basel: Karger 555,556.
Cohen, G. M., Sun, X.-M., Snowden, R. T., Dinsdale, D., and Skilleter, D. N. (1992) Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. Biochem. J. 286, 331–334.
Fadok, V. A., Voelker, D. R., Campbell, P. A., Cohen, J. J., Bratton, D. L., and Henson, P. M. (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148, 2207–2216.
Rodriguez-Tarduchy, G., Collins, M., and Lopez-Rivas, A. (1990) Regulation of apoptosis in interleukin-3 dependent hemopoietic cells by interleukin-3 and calcium ionophores. EMBO J. 9, 2997–3002.
Lennon, S. V., Martin, S. J., and Cotter, T. G. (1991) Dose-dependent induction of apoptosis in tumour cell lines by widely divergent stimuli. Cell Prolif. 24, 203–214.
Martin, S. J. and Cotter, T. G. (1991) Ultraviolet B irradiation of human leukaemia HL-60 cells in vitro induces apoptosis. Int. J. Radiat. Biol. 59, 1001–1016.
Hickman, J. A. (1992) Apoptosis induced by anticancer drugs. Cancer Metastasis Rev. 11, 121–139.
Trauth, B. C., Klas, C., Peters, A. M. J., Matzku, S., Moller, P., Falk, W., Debatin, K.-M., and Krammer, P. H. (1989) Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science 245, 301–305.
McGahon, A. J., Martin, S. V., Bissonnette, R. P., Mahboubi, A., Shi, Y., Mogil, R. J., Nishioka, W. K., and Green, D. R. (1995) The end of the (cell) line: methods for the study of apoptosis in vitro, in Cell Death (Schwartz, I. M. and Osborne B. A., eds.), Academic, San Diego, pp. 153–185.
Martin, S. J., Lennon, S. V., Bonham, A. M., and Cotter, T. G. (1990) Induction of apoptosis (programmed cell death) in human leukaemic HL-60 cells by inhibition of RNA or protein synthesis. J. Immunol. 145, 1859–1867.
Cotter, T. G., Lennon, S. V., Glynn, J. M., and Green, D. R., (1992) Microfilament-disrupting agents prevent formation of apoptotic bodies in tumour cells undergoing apoptosis. Cancer Res. 25, 997–1005.
Swat, W., Ignatowicz, L., and Kieslow, P. (1991) Detection of apoptosis of immature CD4+8+ thymocytes by flow cytometry. J. Immunol. Methods 137, 79–87.
Samali, A. and Cotter, T. G. (1996) Heat shock proteins increase resistance to apoptosis. Exp. Cell Res. 223, 163–170.
Wyllie, A. H., Morris, R. G., Smith, A. L., and Dunlop, D. (1984) Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macro molecular synthesis. J. Pathol. 142, 67–77.
Gorczyca, W., Bruno, S., Darzynkiewicz, R. J., Gong, J., and Darzynkiewicz, Z. (1992) DNA strand breaks occurring during apoptosis: their early detection by terminal deoxynucleotide transferase and nick translation assays and prevention by serine protease inhibitors. Int. J. Oncol. 1, 639–648.
Brown, D. G., Sun, X.-M., and Cohen, G. M. (1993) Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. J. Biol. Chem. 268, 3037–3039.
Cohen, G. M., Sun, X.-M., Fearhead, H., MacFarlane, M., Brown, D. G., Snowden, R. T., and Dinsdale, D. (1994) Formation of large molecular fragments of DNA is a key committed step of apoptosis in thymocytes. J. Immunol. 153, 507–516.
Koopman, G., Reutelingsperger, C. P. M., Kuijten, G. A. M., Keehnen, R. M. J., Pals, S. T., and Van Oers, M. H. J. (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84, 1415–1420.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Samali, A., Cotter, T.G. (1999). Measurement of Cell Death in Culture. In: Jenkins, N. (eds) Animal Cell Biotechnology. Methods in Biotechnology™, vol 8. Humana Press. https://doi.org/10.1385/0-89603-547-6:155
Download citation
DOI: https://doi.org/10.1385/0-89603-547-6:155
Publisher Name: Humana Press
Print ISBN: 978-0-89603-547-8
Online ISBN: 978-1-59259-486-3
eBook Packages: Springer Protocols