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Under sterile conditions, the cells are pelleted by centrifugation at 75g for 5 min at 4°C. 40 Cristofalo, Volker, and Allen b. ). c. ). 3. 5 mL) 4. , steps 13–17, except that serum-free cell growth medium is used. 3. Replicative Life-Span As noted previously, cells in culture exhibit a finite number of replications. At the end of their in vitro life-span substantial cell death occurs; however, a stable population emerges that can exist in a viable, though nondividing, state indefinitely (128).

1989) Growth-factor-inducible gene expression in senescent human fibroblasts. Exp. Gerontol. 24, 515–522. 84. , Croce, M. , and Franceschi, C. (1996) C-Fos/C-Jun expression and AP-1 activation in skin fibroblasts from centenarians. Biochem. Biophys. Res. Commun. 226, 517–523. 85. Millis, A. , McCue, H. , and Martini, H. (1992) Differential expression of metalloproteinase and tissue inhibitor of metalloproteinase genes in aged human fibroblasts. Exp. Cell Res. 201, 373–379. 86. Pignolo, R. , Cristofalo, V.

And Harrison, D. ), Alan Liss, New York, pp. 5–39. 73. Danes, B. S. (1971) Progeria: a cell culture study on aging. J. Clin. Invest. 50, 2000–2003. 74. Schneider, E. L. and Epstein, C. J. (1972) Replication rate and lifespan of cultured fibroblasts in Down’s syndrome. Proc. Soc. Exp. Biol. Med. 141, 1092–1094. 75. Martin, G. , Ogburn, C. , and Sprague, C. A. (1981) Effects of age on cell division capacity, in Aging: A Challenge to Science and Society, vol. , Shock, N. ), Oxford University Press, Oxford, England, pp.

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