Abstract
Mathematical modeling and computational simulations are often used to explain the stochastic nature of cell aging. The models published thus far are based on the molecular mechanisms of telomere biology and how they dictate the dynamics of cell culture proliferation. However, the influence of cell growth conditions on telomere dynamics has been widely overlooked. These conditions include interactions with surrounding cells through contact inhibition, gradual accumulation of non-dividing cells, culture propagation and other cell culture maintenance factors. In order to follow the intrinsic growth dynamics of normal human fibroblasts we employed the fluorescent dye DiI and FACS analysis which can distinguish cells that undergo different numbers of divisions within culture. We observed rapid generation of cell subpopulations undergoing from 0 to 9 divisions within growing cultures at each passage analyzed. These large differences in number of divisions among individual cells guarantee a strong impact on generation of telomere length heterogeneity in normal cell cultures and suggest that culture conditions should be included in future modeling of cell senescence.
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References
Arino O, Kimmel M, Webb GF (1995) Mathematical modeling of the loss of telomere sequences. J Theor Biol 177:45–57
Arkus N (2005) A mathematical model of cellular apoptosis and senescence through the dynamics of telomere loss. J Theor Biol 235:13–32
Bourgeron T, Xu Z, Doumic M, Teresa Teixeira M (2015) The asymmetry of telomere replication contributes to replicative senescence heterogeneity. Sci Rep 5:15326
Calado RT, Young NS (2009) Telomere diseases. N Engl J Med 361:2353–2365
Calado R, Young N (2012) Telomeres in disease. F1000 Med Rep 4:8
Counter CM, Avilion AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB, Bacchetti S (1992) Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:1921–1929
Cukusic Kalajzic A, Škrobot Vidaček N, Huzak M, Ivankovic M, Rubelj I (2014) Telomere Q-PNA-FISH—reliable results from stochastic signals. PLoS ONE 9:e92559
de Lange T (2002) Protection of mammalian telomeres. Oncogene 21:532–540
de Lange T, Zhu X-D, Küster B, Mann M, Petrini JHJ (2000) Cell-cycle-regulated association of RAD50/MRE11/NBS1 with TRF2 and human telomeres. Nat Genet 25:347–352
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O et al (1995) A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 92:9363–9367
Ferenac M, Polancec D, Huzak M, Pereira-Smith O, Rubelj I (2005) Early-senescing human skin fibroblasts do not demonstrate accelerated telomere shortening. J Gerontol A Biol Sci Med Sci 60:820–829
Griffith JD, Comeau L, Rosenfield S, Stansel RM, Bianchi A, Moss H, de Lange T (1999) Mammalian telomeres end in a large duplex loop. Cell 97:503–514
Harley CB, Futcher AB, Greider CW (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460
Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636
Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 25:585–621
Hemann MT, Strong MA, Hao LY, Greider CW (2001) The shortest telomere, not average telomere length, is critical for cell viability and chromosome stability. Cell 107:67–77
Levy MZ, Allsopp RC, Futcher AB, Greider CW, Harley CB (1992) Telomere end-replication problem and cell aging. J Mol Biol 225:951–960
Nelson G, Wordsworth J, Wang C, Jurk D, Lawless C, Martin-Ruiz C, von Zglinicki T (2012) A senescent cell bystander effect: senescence-induced senescence. Aging Cell 11:345–349
Olofsson P, Bertuch AA (2010) Modeling growth and telomere dynamics in Saccharomyces cerevisiae. J Theor Biol 263:353–359
Olofsson P, Kimmel M (1999) Stochastic models of telomere shortening. Math Biosci 158:75–92
Portugal RD, Land MGP, Svaiter BF (2008) A computational model for telomere-dependent cell-replicative aging. Biosystems 91:262–267
Proctor CJ, Kirkwood TBL (2002) Modelling telomere shortening and the role of oxidative stress. Mech Ageing Dev 123:351–363
Proctor CJ, Kirkwood TBL (2003) Modelling cellular senescence as a result of telomere state. Aging Cell 2:151–157
Rodier F, Campisi J (2011) Four faces of cellular senescence. J Cell Biol 192:547–556
Rubelj I, Vondracek Z (1999) Stochastic mechanism of cellular aging—abrupt telomere shortening as a model for stochastic nature of cellular aging. J Theor Biol 197:425–438
Rubelj I, Huzak M, Brdar B (2000) Sudden senescence syndrome plays a major role in cell culture proliferation. Mech Ageing Dev 112:233–241
Rubelj I, Huzak M, Brdar B, Pereira-Smith OM (2002) A single-stage mechanism controls replicative senescence through Sudden Senescence Syndrome. Biogerontology 3:213–222
Shay JW, Wright WE (2001) Telomeres and telomerase: implications for cancer and aging. Radiat Res 155:188–193
Tan Z (1999) Intramitotic and intraclonal variation in proliferative potential of human diploid cells: explained by telomere shortening. J Theor Biol 198:259–268
Vidaček NŠ, Ćukušić A, Ivanković M, Fulgosi H, Huzak M, Smith JR, Rubelj I (2010) Abrupt telomere shortening in normal human fibroblasts. Exp Gerontol 45:235–242
von Zglinicki T (2002) Oxidative stress shortens telomeres. Trends Biochem Sci 27:339–344
Wu P, Takai H, de Lange T (2012) Telomeric 3′ overhangs derive from resection by Exo1 and Apollo and fill-in by POT1b-associated CST. Cell 150:39–52
Xu Z, Duc KD, Holcman D, Teixeira MT (2013) The length of the shortest telomere as the major determinant of the onset of replicative senescence. Genetics 194:847–857
Acknowledgements
We thank Mary Sopta for critically reading and editing the manuscript, Ela Kosor and Alenka Gagro for their participation in FACS analysis. We also thank Milena Ivanković, Marina Ferenac Kiš, Maja Matulić and Andrea Ćukušić Kalajžić for valuable discussions and practical assistance during the course of the experiments. This work was supported by Zaklada Adris.
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Supplementary Fig. 7
[3H]thymidine labelling and SA-β Gal staining of NF fibroblasts at PD 10. a) Senescent cells are enlarged, they do not incorporate radioactivity and show strong SA-β Gal staining (purple arrow); young cells are smaller, 3H-T+ and do not show SA-β Gal staining (yellow arrow) b) Most cells were dividing and total of 97.25% incorporated radioactivity, among which 11.76% also showed traces of SA-β-Gal staining. > 1000 cells were counted for statistics (TIFF 1053 kb)
Supplementary Fig. 8
[3H]thymidine labelling index and SA-β Gal staining of NF fibroblasts at PDs 10 and 15. > 1000 cells were counted for statistics (TIFF 126 kb)
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Nanić, L., Vidaček, N.Š., Ravlić, S. et al. Mutual interactions between telomere heterogeneity and cell culture growth dynamics shape stochasticity of cell aging. Biogerontology 19, 23–31 (2018). https://doi.org/10.1007/s10522-017-9736-2
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DOI: https://doi.org/10.1007/s10522-017-9736-2