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104. Lapiere, C. M. (1988) Aging of fibroblasts, collagen and the dermis, in Cutaneous Aging (Kligman, A. M. ), University of Tokyo Press, Tokyo, 47–60. 105. Choi, A. M. , Olsen, D. , Cook, K. , Deamond, S. , and Bruce, S. A. (1992) Differential extracellular matrix gene expression by fibroblasts during their proliferative life span in vitro and senescence. J. Cell. Physiol. 151, 147–155. 106. , Levine, E. , and Eagle, H. (1970) Morphologic changes accompanying senescence of cultured human diploid cells.

Cell 8, 2157–2169. 48 Cristofalo, Volker, and Allen 61. , Lin, A. , McCurrach, M. , and Lowe, S. W. (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16ink4a. Cell 88, 593–602. 62. Rohme, D. (1981) Evidence for a relationship between longevity of mammalian species and life spans of normal fibroblasts in vitro and erythrocytes in vivo. Proc. Natl. Acad. Sci. USA 78, 5009–5013. 63. Hayflick, L. (1965) The limited in vitro lifetime of human diploid strains.

Remove the coverslips from the NaOH solution and rinsed at least three times in deionized water. 8. Remove the coverslips from the rack and allow to air-dry on lint-free disposable wipes. 9. When completely dry, bake the coverslips for 3 h at 180°C for sterilization. 2. Cell Slides 1. In a laminar flow hood, under sterile conditions, cells are harvested and counted in the usual manner. 2. ). If using coverslips use sterile forceps to arrange them in the dish so that they do not overlap one another.

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