Severino J, Allen R G, Balin S, Balin A, Cristofalo V J
Center for Gerontological Research, MCP-Hahnemann University, Philadelphia, Pennsylvania 19129, USA.
Exp Cell Res. 2000 May 25;257(1):162-71. doi: 10.1006/excr.2000.4875.
Cytochemically detectable beta-galactosidase (beta-gal) at pH 6.0 has been reported to increase during the replicative senescence of fibroblast cultures and has been used widely as a marker of cellular senescence in vivo and in vitro. In this study, we have characterized changes in senescence-associated (SA) beta-gal staining in early and late passage cultures, cultures established from donors of different ages, virally immortalized cells, and tissue slices obtained from donors of different ages. The effects of different culture conditions were also examined. While we confirm the previous report that SA beta-gal staining increased in low-density cultures of proliferatively senescent cells, we were unable to demonstrate that it is a specific marker for aging in vitro. Cultures established from donors of different ages stained for SA beta-gal activity as a function of in vitro replicative age, not donor age. We also failed to observe any differences in SA beta-gal staining in skin cells in situ as a marker of aging in vivo. The level of cytochemically detectable SA beta-gal was elevated in confluent nontransformed fibroblast cultures, in immortal fibroblast cultures that had reached a high cell density, and in low-density, young, normal cultures oxidatively challenged by treatment with H2O2. Although we clearly demonstrate that SA beta-gal staining in cells is increased under a variety of different conditions, the interpretation of increased staining remains unclear, as does the question of whether the same mechanisms are responsible for the increased SA beta-gal staining observed in senescent cells and changes observed in cells under other conditions.
据报道，在pH 6.0时细胞化学可检测的β-半乳糖苷酶（β-gal）在成纤维细胞培养物的复制性衰老过程中会增加，并已在体内和体外广泛用作细胞衰老的标志物。在本研究中，我们对早期和晚期传代培养物、从不同年龄供体建立的培养物、病毒永生化细胞以及从不同年龄供体获得的组织切片中衰老相关（SA）β-gal染色的变化进行了表征。还研究了不同培养条件的影响。虽然我们证实了先前的报道，即SA β-gal染色在增殖性衰老细胞的低密度培养物中增加，但我们无法证明它是体外衰老的特异性标志物。从不同年龄供体建立的培养物中SA β-gal活性的染色是体外复制年龄的函数，而不是供体年龄的函数。我们也未能观察到原位皮肤细胞中SA β-gal染色作为体内衰老标志物的任何差异。在汇合的未转化成纤维细胞培养物、达到高细胞密度的永生化成纤维细胞培养物以及用H2O2处理进行氧化挑战的低密度年轻正常培养物中，细胞化学可检测的SA β-gal水平升高。虽然我们清楚地证明了在各种不同条件下细胞中SA β-gal染色增加，但染色增加的解释仍然不清楚，衰老细胞中观察到的SA β-gal染色增加以及其他条件下细胞中观察到的变化是否由相同机制引起的问题也不清楚。
