Abstract
Senescence-associated β-galactosidase (SA-β-Gal) is a widely used marker of senescent cells in vitro and in vivo. In this report, young and senescent human peritoneal mesothelial cells (HPMCs) and fragments of the omentum, from which these cells were isolated, were subjected to simultaneous examination of SA-β-Gal using two methods, i.e. cytochemical and fluorescent methods. The results obtained were confronted with the cumulative number of population doublings (CPD) and the calendar age of the tissue donor. The study showed that senescence of HPMCs proceeds with either an increased percentage of SA-β-Gal-positive cells or increased enzyme activity. Cytochemical SA-β-Gal staining in early-passage cultures negatively correlated with CPD values but not with donor age in both cell cultures and omentum specimens. Conversely, SA-β-Gal activity measured with the fluorescence method rose in proportion to the calendar age of the donor either in early-passage cultures or in primary cell isolates from omental tissue. At the same time it was not related to the CPD values. These findings may suggest that with respect to at least peritoneal mesothelial cells, the cytochemical and fluorescent methods of SA-β-Gal detection, though complementary, are informative for different levels of aging, i.e. the cytochemical approach for senescence in vitro and the fluorescence-based technique for organismal aging in vivo.
Highlights
Replicative senescence refers to permanent growth arrest of normal somatic cells elicited by their passage through a fixed number of divisions
There is a wealth of evidence that replicative senescence of human peritoneal mesothelial cells (HPMCs) contributes to organismal, or at least omental, tissue aging
These include an age-dependent pattern of oxidative DNA damage intensification (Ksiazek et al 2008), an accumulation of senescent cells in the omentum in vivo (Ksiazek et al 2008b), and an inverse relationship between donor age and cell expandability in vitro (Ksiazek et al 2007b)
Summary
Patrycja Sosinska • Justyna Mikuła-Pietrasik • Monika Ryz_ek • Eryk Naumowicz • Krzysztof Ksiaz_ek Received: 3 February 2014 / Accepted: 12 May 2014 / Published online: 31 May 2014 Ó The Author(s) 2014. This article is published with open access at Springerlink.com
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