Rapid flow cytometric method for measuring senescence associated β-galactosidase activity in human fibroblasts

Abstract

Senescence associated-β-galactosidase (SA-β-gal) activity is a widely used marker for cellular senenescence. SA-β-gal activity is routinely detected cytochemically, manually discriminating negative from positive cells. This method is time-consuming, subjective and therefore prone to operator-error. We aimed to optimize a flow cytometric method described by other workers using endothelial cells to better differentiate between populations of fibroblasts in degrees of SA-β-gal activity. Skin fibroblasts were isolated from young (mean age ± SD: 25.5 ± 1.8) and very old (age 90.2 ± 0.3) subjects. Different pH modulators were tested for toxicity. To induce stress-induced senescence, fibroblasts we..