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The limitations and validities of senescence associated-β-galactosidase activity as an aging marker for human foreskin fibroblast Hs68 cells
Abstract
The senescence associated-beta-galactosidase (SA-betaG) assay has become one of the most commonly used markers of cell-aging. However, the reliability of the assay is questionable because the enzyme is a non-specific marker for cell-aging. In this study, we found that the SA-betaG activity increased with cell age as well as in confluent quiescent cells or cells under serum starvation, and in cells treated with H2O2. Importantly, we found that SA-betaG activity was irreversibly increased in the senescent cells or H2O2-teated cells, but was reversible in quiescent cells induced by serum starvation or confluence. Using fluorescein di-beta-d-galactopyranoside (FDG) method for SA-betaG detection, we showed that senescent human foreskin fibroblast Hs68 cells did not express a specific enzyme that has a maximal activity at pH 6.0. In the pH profile of the cellular betaG activity in senescent Hs68 cells, only a single peak was found (with maximum at pH 4.6), and no addition peak was found at or around pH 6.0 that could be attributed to the SA-betaG activity. These results support the contention that SA-betaG is the lysosomal betaG that is detectable at suboptimal pH (i.e. pH 6.0) and demonstrate that cell-aging is not the only factor that can increase SA-betaG activity, rendering SA-betaG activity unspecific for cell-aging. Thus, the assay for cell-aging is only reliable when these confounding factors are controlled or excluded.
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Additional stress conditions such as long-term confluence and prolonged serum starvation can trigger SA-β-Gal activity.
“Moreover, additional stress conditions such as long-term confluence and prolonged serum starvation can trigger SA-β-Gal activity23.”
From Algorithmic assessment of cellular senescence in experimental and clinical specimens (2021) · cites this paper as [23]
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Metadata
- Authors
- YANG, N., & HU, M.
- Affiliations
- National Chung Hsing University (TW)
- Corresponding author
- HU, M
- Venue
- Experimental Gerontology
- Publisher
- Elsevier BV
- Published
- 2005-10
- Volume
- 40
- Issue
- 10
- Pages
- 813-819
- ISSN
- 0531-5565 (ISSN-L)
- OpenAlex
- W2020595943
- Cited by
- 245
- References
- 16
- Citations by year
- 2026: 12 · 2025: 19 · 2024: 22 · 2023: 18 · 2022: 19 · 2021: 25 · 2020: 11 · 2019: 14 · 2018: 10 · 2017: 8 · 2016: 13 · 2015: 9 · 2014: 11 · 2013: 9 · 2012: 6
- FWCI
- 3.21
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Experimental Gerontology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Telomeres, Telomerase, and Senescence
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Antioxidants, Aging, Portulaca oleracea
- Keywords
- senescence-associated β-galactosidase, cellular senescence, aging markers, lysosomal beta-galactosidase, pH profile, serum deprivation, hydrogen peroxide, quiescent cells, assay specificity
- MeSH terms
- Aged, beta-Galactosidase, Cell Line, Fibroblasts, Histocytochemistry, Humans, Male, Models, Biological, Skin, Biomarkers, Cellular Senescence
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