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KMID : 0385220040140020031
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Korean Journal of Gerontology
2004 Volume.14 No. 2 p.31 ~ p.38
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Screening of Anti-aging Materials using Cell Culture of Human Fibroblasts
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Kim Hyun-Jung
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Abstract
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Since aging is a complex and life time-consuming process, it is hard to measure anti-aging activity of materials. Longevity could be measured using laboratory animals, rat and mouse but it takes hundreds of animals at least for 3 - 4 years. When numbers of materials to test for longevity are many, it is almost impossible. Therefore, we exploit the cellular replicative senescence of human fibroblast to develop an easy in vitro screening method of anti-aging materials, since the replicative senescence of human diploid fibroblast reported to mimic in vivo aging in various points. A human embryonic fibroblast line which was established from an abortus by Hallym University Hospital, was obtained and used. Growth curve of this line was prepared and this line reached replicative senescence at 66 PD (population doubling). The levels of p21 protein and mRNA were increased and SA-beta-GAL (senescence-associated beta galactosidase) was activated at replicative senescence as expected. When PBN (a-phenyl-N-butyl nitrone) was added to cell culture at 40 PD as an anti-aging material, it extended the maximum doubling number of the fibroblast line by 8-10 PD and reduced SA-beta-GAL staining by 40% compared to untreated cells. Therefore, by measuring simply maximum population doubling of human fibroblast line and by calculating the percentage of SA-beta-GAL positive cells at 60 PD after adding materials various concentration in 40 to 50 PD, anti-aging activity of materials could be evaluated through comparing to untreated control or known materials. The procedure is apparently a very useful screening method for anti-aging activity since it does not take that much time and cost.
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KEYWORD
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human fibroblast, replicative senescence, anti-aging materials, screening
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