By Yvonne A. Barnett
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Extra resources for Aging Methods and Protocols (Methods in Molecular Medicine)
Emulsion: Kodak NTB-2 emulsion is purchased in a lightproof container. The emulsion is stored at 4°C (see Note 6). 4. Developer and Fixer a. Kodak D-19 developer is purchased in packets that make 1 gal when reconstituted. The entire packet is used at one time and the solution is stored in a brown bottle in the dark. The developer remains useable for 1–3 mo. When the developer turns yellow, it is discarded. b. Acid fixer is made and stored in the same manner as the D-19 developer. 3. 1. Cell Propagation in Serum-Supplemented Medium Cells may be grown in a variety of culture vessels (see Note 7).
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). Furthermore, small subpopulations of cells may retain some growth capacity even after the vast majority of cells in a culture are no longer able to divide.
Enlow, B. , Saati, S. , Tonkin, L. , Funk, W. , and Villeponteau, B. (1995) Cataloging altered gene expression in yong and senescent cells using enhanced differential display. Nucleic Acids Res. 23, 3244–3251. 115. , Vinci, J. , Millis, A. J. , and Baglioni, C. (1993) Expression of interleukin-1α and β in early passage fibroblasts from aging individuals. Exp. Gerontol. 28, 505–513. 116. Karlsson, C. and Paulsson, Y. (1994) Age-related induction of platelet-derived growth factor A-chain mRNA in normal human fibroblasts.