Summary
Immortal cells perpetuate the rises and falls of proliferation that are progressively damped in mortal long-term cultured cells. For immortal rat hepatoma Fao cells, similar waves of proliferation occurred about every 3–4 wk. Under the same conditions, embryonic human fibroblasts and transformed but not immortalized embryonic fibroblasts display similarly recurring proliferation waves that progressively decrease in amplitude until senescence of the lines. In addition, strains of diploid normal human skin fibroblasts cultured under different culture conditions display a similar time-pattern of proliferation. Although the amplitude and baseline of these fluctuations are characteristic for each cell line, a common point was marked slow down in proliferation after every sequence of about 25 population doublings for all cells. Renewed proliferation waves of Fao cells allow about 22–23 additional population doublings each. Normal embryonic fibroblasts culture and its transformed counterpart accumulate about 30 and 60 population doublings, respectively, before senescence. Normal fibroblast strains accumulate about 25 population doublings over their entire life spans. This halt in proliferation after every stretch of about 25 population doublings may correspond to a structural or functional stop following attrition of telomeric DNA. This putative stop may be bypassed once in transformed embryonic cells and repetitively in immortal cells. In support of this hypothesis, we observed rapid telomere shortening, in two steps, during divisions of mortal embryonic cells, and maintenance of long telomeres in immortal Fao cells, which may indicate episodic repair of telomeres. Alternatively, such maintenance of long telomeres may reflect survival and successive clonal growth of rare cells with long telomeres. We suggest that the balance between telomere attribution and repair processes regulates the waves of proliferation.
Similar content being viewed by others
References
Allsopp, R. C.; Vaziri, H.; Patterson, C., et al. Telomere length predicts replicative capacity of human fibroblasts. Proc. Natl. Acad. Sci. USA 89:10114–10118; 1992.
Bell, E.; Marek, L. F.; Levinstone, D. S., et al. Loss of division potential in vitro: aging or differentiation? Science 202:1158–1163; 1978.
Benoît, C.; Chambon, P. In vivo requirements of the SV40 early promoter region. Nature 290:304–309; 1981.
Chikappa, G.; Borner, G.; Burlington, H., et al. Periodic oscillation of blood leukocytes, platelets, and reticulocytes, in a patient with chronic myelocytic leukemia. Blood 47:1023–1030; 1976.
Counter, C. M.; Avilion, A. A.; LeFeuvre, C. E., et al. Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J. 11:1921–1929; 1992.
Counter, C. M.; Hirte, H. W.; Bacchetti, S., et al. Telomerase activity in human ovarian carcinoma. Proc. Natl. Acad. Sci. USA 91:2900–2904; 1994.
Cristofalo, V. J.; Sharf, B. B. Cellular senescence and DNA synthesis. Exp. Cell Res. 76:419–427; 1973.
Deschatrette, J.; Weiss, M. C. Characterization of differentiated and dedifferentiated clones of a rat hepatoma. Biochimie 56:1603–1611; 1974.
Foulds, L. Mammary tumours in hybrid mice: growth and progression of spontaneous tumors. Br. J. Cancer 3:345–375; 1949.
Gatti, R. A.; Robinson, W. A.; Deinard, A. S., et al. Cyclic leukocytosis in chronic myelogenous leukemia: new perspectives on pathogenesis and therapy. Blood 41:771–782; 1973.
Harley, C. B.; Futcher, A. B.; Greider, C. W. Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460; 1990.
Harley, C. B.; Vaziri, H.; Counter, C., et al. The telomere hypothesis of cellular aging. Exp. Gerontol. 4:375–382; 1992.
Hayflick, L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37:611–635; 1964.
Holt, S. E.; Wright, W. E.; Shay, J. W. Regulation of telomerase activity in immortal cell lines. Mol. Cell. Biol. 6:2932–2939; 1996.
Juckett, D. A. Cellular aging (the Hayflick limit) and species longevity: a unification model based on clonal succession. Mech. Ageing Dev. 38:49–71; 1987.
Karatza, C.; Stein, W. D.; Shall, S. Kinetics of in vitro ageing of mouse embryo fibroblasts. J. Cell Sci. 65:163–175; 1984.
Kennedy, B. J. Cyclic leukocyte oscillations in chronic myelogenous leukemia during hydroxyurea therapy. Blood 35:751–760; 1970.
Kim, N. W.; Piatyszek, M. A.; Prowse, K. R., et al. Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015; 1994.
Levy, M. Z.; Allsopp, R. C.; Futcher, A. B., et al. Telomere end-replication problem and cell aging. J. Mol. Biol. 225:951–960; 1992.
Maigné, J.; Ng, K. H.; Meunier-Rotival, M., et al. Correlation between reversion of a dedifferentiated rat hepatoma line and the recovery of tumorigenicity. Cancer Res. 48:3258–3264; 1988.
Martin, G. M.; Sprague, C. A.; Epstein, C. J. Repliative life-span of cultivated human cells. Effects of donor’s age, tissue, and genotype. Lab. Invest. 23:86–92; 1970.
McEachern, M. J.; Blackburn, E. H. Runaway telomere elongation caused by telomerase RNA gene mutations. Nature 376:403–409; 1995.
Miles, C. P. Prolonged culture of diploid human cells. Cancer Res. 24:1070–1081; 1964.
Moore, J. V.; Rowley, R.; Hopkins, H. A., et al. Cyclophosphamide as an adjuvant to X-rays in treatment of a radioresistant solid tumor of the rat, hepatoma H-4-II-E. Int. J. Radiat. Oncol. Biol. Phys. 5:1471–1474; 1979.
Pitot, H. C.; Peraino, C.; Morse, P. A., et al. Hepatoma in tissue culture compared with adapting liver in vivo. Natl. Cancer Inst. Monog. 13:229–242; 1964.
Prowse, K. R.; Greider, C. W. Developmental and tissue-specific regulation of mouse telomerase and telomere length. Proc. Natl. Acad. Sci. USA 92:4818–4822; 1995.
Reuber, M. D. A transplantable bile-secreting hepatocellular carcinoma in the rat. J. Natl. Cancer Inst. 26:891–897; 1961.
Schächter, F.; Boucher, N.; Lesueur-Ginot, L., et al. Sénescence cellulaire et survie des lymphocytes T. Cellular senescence and survival of T lymphocytes. Comptes Rendus Acad. Sci. Paris 318:56572; 1995.
Shay, J. W.; Brasiskyte, D.; Ouellette, M., et al. Analysis of telomerase and telomeres. Methods Mol. Genet. 5:263–280; 1994.
Smith, J. R.; Hayflick, L. Variation in the lifespan of clones derived from human diploid cell strains. J. Cell Biol. 62:48–53; 1974.
Speer, J. F.; Petrosky, V. E.; Retsky, M. W., et al. A stochastic numerical model of breast cancer growth that simulates clinical data. Cancer Res. 44:4124–4130; 1984.
Squartini, F. Strain differences in growth of mammary tumors. J. Natl. Cancer Inst. 26:813–828; 1961.
Tommerup, H.; Dousmanis, A.; de Lange, T. Unusual chromatin in human telomeres. Mol. Cell. Biol. 9:5777–5785; 1994.
Wolfrom, C.; Raynaud, N.; Maigné, J., et al. Periodic fluctuations in proliferation of SV-40 transformed human skin fibroblast lines with prolonged lifespan. Cell Biol. Toxicol. 10:247–254; 1994.
Zhong, Z.; Shiue, L.; Kaplan, S., et al. A mammalian factor that binds telomere TTAGGG repeats in vitro. Mol. Cell. Biol. 12:4834–4843; 1992.
Author information
Authors and Affiliations
Additional information
Equal contributors to these studies.
Rights and permissions
About this article
Cite this article
Maigné, J., Deschatrette, J., Sarrazin, S. et al. The time-pattern of rises and falls in proliferation fades with senescence of mortal lines and is perpetuated in immortal rat hepatoma fao cell line. In Vitro Cell.Dev.Biol.-Animal 34, 163–169 (1998). https://doi.org/10.1007/s11626-998-0100-3
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11626-998-0100-3