The International Journal of Biochemistry & Cell Biology
ReviewFrom the Hayflick mosaic to the mosaics of ageing.: Role of stress-induced premature senescence in human ageing
Section snippets
The Hayflick limit today
In 1961 Hayflick and Moorhead reported that human diploid fibroblasts (HDFs) divide a finite number of times. Hayflick called ‘phase I’ the primary culture, ‘phase II’ the many cumulative populations doublings (CPDs) of luxuriant growth after the primary culture, and ‘phase III’ the subsequent growth arrest [1], [2]. In 1974 ‘phase III’ was termed ‘the Hayflick limit’ [3]. The phrase ‘replicative senescence’ is now widely used. Medline refers to more than 6100 papers with ‘replicative
Effects of culture conditions on senescence
The deregulation of the mitotic machinery of HDFs during in vivo ageing of middle-aged and late-aged donors was analysed with DNA μarrays. Many changes were found in the expression level of genes necessary for the cell cycle to proceed [14]. Most of these genes, however, are different from those undergoing expression changes in in vitro replicatively senescent HDFs. This suggests that in vivo senescence is somewhat different from in vitro replicative senescence.
When cultured under 3% O2, i.e.
The Hayflick mosaic
In vivo the proliferative capacity of HDFs is never completely exhausted. HDFs of centenarians are still able to divide in vitro, sometimes for a number of CPDs that renders them undiscriminable from explants of HDFs of young donors [43]. Stem cells are present in the connective tissues of dermis and skeletal muscle derived from geriatric humans. These cells contain lineage-committed myogenic, adipogenic, chondrogenic, and osteogenic progenitor stem cells as well as lineage-uncommitted
The mosaics of ageing
The narrowest definition of senescence is irreversible growth arrest triggered by telomere shortening, which counts cell generations [11] (definition 1). Other authors enlarged this definition to a functional definition encompassing all kinds of irreversible arrests of proliferative cell types including that induced by damaging agents (definition 2) [6]. Irreversible growth arrest of proliferative cell types induced by damaging agents may be called stress-induced senescence-like phenotype,
SIPS in vitro
Many proliferative cell types (lung and skin HDFs, human melanocytes, endothelial cells, human retinal pigment epithelial cells, human erythroleukemia cells, etc.) exposed to subcytotoxic stress (UV, organic peroxides, H2O2, ethanol, mitomycin C, hyperoxia, γ-irradiations, homocysteine, hydroxyurea, tert-butylhydroperoxide (t-BHP) etc.) undergo SIPS. SIPS can be defined as the long term effects of subcytotoxic stress on proliferative cell types, including irreversible growth arrest of (a
Growth arrest
HDFs in H2O2-induced SIPS cannot launch a mitogenic response when stimulated with serum or usual growth factors [68]. Most of HDFs in H2O2-induced SIPS are growth arrested in the G1 phase of the cell cycle [65]. Hyperoxia under 40% O2 also leads to growth arrest of HDFs in the G1 phase [69].
The proportion of HDFs positive for SA β-gal activity correlates with CPDs. It also increases in SIPS, induced by tert-butylhydroperoxide (t-BHP) and H2O2 (for a review: [64]). SA β-gal activity is rather a
Is telomere shortening involved in SIPS?
WI-38 HDFs kept under 40% O2 for 3 CPDs undergo SIPS. An accelerated telomere restriction fragment (TRF) shortening (500 bp/PD) is observed [69]. Forty percent O2 induces single-strand breaks and accelerated TRF shortening of human retinal pigment epithelial cells [89]. WI-38 HDFs undergo accelerated TRF (490 bp/stress) and irreversible growth arrest after four exposures to subcytotoxic t-BHP stress, with a stress at every 2 CPDs. After these stresses, the cells stop proliferating. The control
Hayflick mosaic or mosaics of ageing?
From definition 1 of senescence, the fact that HDFs can endure more PDs at physiological low O2 partial pressures decreases the probabilities to find senescent cells based on the end-replication problem in vivo. Theoretically, starting from the two first telomerase-negative cells that appear during in vivo differentiation, and that will become fibroblasts (in the case of symmetric divisions) 250 cells (>1015 cells) must be produced before the first telomere-dependent replicatively senescent
Conclusions
The models of SIPS are already used in toxicology to seek possible long term effects of molecules in R&D. One can also detect possible anti-ageing effects of molecules in human cells in SIPS. Automation of the models of SIPS will lighten the budgetary and ethical burden of in vivo tests [120]. These systems give more useful mechanistic information compared to the information gained when using lower invertebrate animals as toxicological models.
The appearance of SIPS could be due to exacerbated
Acknowledgements
O. Toussaint is a research associate and F. Chainiaux a research assistant of the FNRS, Belgium. J.-F. Dierick, C. Frippiat, P. Dumont are FRIA fellows, Belgium. J. P. Magalhaes thanks FCT, Portugal. We thank the European Union, 5th Framework Programme, Quality of Life, R&D, ‘Protage’ (QLK6-CT-1999-02193) and ‘Functionage’ (QLK6-CT-2001-00310) and the Région Wallonne Project ‘Modelage’.
References (142)
The limited in vitro lifetime of human diploid cell strains
Exp. Cell Res.
(1965)- et al.
The serial cultivation of human diploid cell strains
Exp. Cell Res.
(1961) - et al.
Microarray analysis of replicative senescence
Curr. Biol.
(1999) - et al.
Putting the stress on senescence
Curr. Opin. Cell Biol.
(2001) - et al.
The shortest telomere, not average telomere length, is critical for cell viability and chromosome stability
Cell
(2001) - et al.
Accumulation of short telomeres in human fibroblasts prior to replicative senescence
Exp. Cell Res.
(2000) - et al.
Cellular senescence as a tumor-protection mechanism: the essential role of counting
Curr. Opin. Genet. Dev.
(2001) - et al.
BJ fibroblasts display high antioxidant capacity and slow telomere shortening independent of hTERT transfection
Free Radic. Biol. Med.
(2001) - et al.
Positive correlation between mammalian life span and cellular resistance to stress
Free Radic. Biol. Med.
(1999) - et al.
Atmospheric oxygen accelerates the induction of a post-mitotic phenotype in human dermal fibroblasts: the key protective role of glutathione
Differentiation
(2000)
Slowing down ageing of cultured embryonal chick chondrocytes by maintenance under lowered oxygen tension
Mech. Ageing Dev.
Importance of a threshold for error accumulation in cell degenerative processes. I. Modulation of the threshold in a model of free radical-induced cell degeneration
Mech. Ageing Dev.
Effect of oxygen on the growth of human epidermal keratinocytes
J. Invest. Dermatol.
Growth-related lipid peroxidation in tumour microsomal membranes and mitochondria
Biochim. Biophys. Acta
Aging as a multi-step process characterized by a lowering of entropy production leading the cell to a sequence of defined stages
Mech. Ageing Dev.
Influence of oxygen tension, pro-oxidants and antioxidants on the formation of lipid peroxidation products (lipofuscin) in individual cultivated human glial cells
Mech. Ageing Dev.
Alterations in mitochondrial membrane fluidity by lipid peroxidation products
Free Radic. Biol. Med.
Effects of the lipidperoxidation product 4-hydroxynonenal and related aldehydes on proliferation and viability of cultured Ehrlich ascites tumor cells
Biochem. Pharmacol.
Cytotoxicity of linoleic acid peroxide, malondialdehyde and 4- hydroxynonenal towards human fibroblasts
Toxicology
Genetic and epigenetic changes in human epithelial cells immortalized by telomerase
Am. J. Pathol.
Growth properties and growth factor responsiveness in skin fibroblasts from centenarians
Biochem. Biophys. Res. Commun.
Replicative mosaicism might explain the seeming contradictions in the telomere theory of aging
Mech. Ageing Dev.
Induction of replicative senescence biomarkers by sublethal oxidative stresses in normal human fibroblast
Free Radic. Biol. Med.
Quantification of wild-type mitochondrial DNA and its 4.8 kb deletion in rat organs
Biochem. Biophys. Res. Commun.
Mitochondrial DNA deletions in human cardiac tissue show a gross mosaic distribution
Biochem. Biophys. Res. Commun.
Attenuated expression of 70 kDa heat shock protein in WI-38 human fibroblasts during aging in vitro
Exp. Cell Res.
Age-dependant decrease in the heat-inducible DNA sequence-specific binding activity in human diploid fibroblasts
J. Biol. Chem.
Age-related changes of heat shock protein gene transcription in human peripheral blood mononuclear cells
Biochem. Biophys. Res. Commun.
Cellular senescence: mitotic clock or culture shock?
Cell
Stress-induced premature senescence (SIPS) of human diploid fibroblasts and melanocytes
Exp. Gerontol.
Sublethal H2O2 stress triggers a release of TGF-β 1 which induce biomarkers of cellular senescence of human diploid fibroblasts
J. Biol. Chem.
DNA synthesis and Fos and Jun protein expression in mitotic and postmitotic WI-38 fibroblasts in vitro
Exp. Cell Res.
Inhibitors of cyclin-dependent kinases induce features of replicative senescence in early passage human diploid fibroblasts
Curr. Biol.
Isolation and identification of psoralen plus ultraviolet A (PUVA)- induced genes in human dermal fibroblasts by polymerase chain reaction- based subtractive hybridization
J. Invest. Dermatol.
Permanent cell cycle arrest in asynchronously proliferating normal human fibroblasts treated with doxorubicin or etoposide but not camptothecin
Biochem. Pharmacol.
Activation of an H2O2-generating NADH oxidase in human lung fibroblasts by transforming growth factor β 1
J. Biol. Chem.
SPARC regulates the expression of collagen type I and transforming growth factor-β1 in mesanglial cells
J. Biol. Chem.
Clusterin/apolipoprotein J is a novel biomarker of senescence that does not affect the proliferative capacity of human diploid fibroblasts
FEBS Lett.
Growth kinetics rather than stress cause accelerated telomere shortening in cultures of human diploid fibroblasts in oxidative stress-induced premature senescence
FEBS Lett.
Uncoupling the senescent phenotype from telomere shortening in hydrogen peroxide-treated fibroblasts
Exp. Cell Res.
Accumulation of single-strand breaks is the major cause of telomere shortening in human fibroblasts
Free Radic. Biol. Med.
Age-dependent telomere shortening is slowed down by enrichment of intracellular Vitamin C via suppression of oxidative stress
Life Sci.
Homocysteine accelerates endothelial cell senescence
FEBS Lett.
Telomere shortening during aging of human osteoblasts in vitro and leukocytes in vivo: lack of excessive telomere loss in osteoporotic patients
Mech. Ageing Dev.
Telomere reduction in human liver tissues with age and chronic inflammation
Exp. Cell Res.
Accelerated telomere shortening in fibroblasts after extended periods of confluency
Free Radic. Biol. Med.
Hayflick, his limit, and cellular ageing
Nat. Rev. Mol. Cell Biol.
Age-dependent modifications of gene expression in human fibroblasts
Crit. Rev. Eukaryot. Gene Expr.
Relationship between donor age and the replicative life spans of human cells in culture: a re-evaluation
Proc. Natl. Acad. Sci. U.S.A.
Cited by (78)
Mimicking Tumor Cell Heterogeneity of Colorectal Cancer in a Patient-derived Organoid-Fibroblast Model
2023, Cellular and Molecular Gastroenterology and HepatologyRole of Prion protein in premature senescence of human fibroblasts
2018, Mechanisms of Ageing and DevelopmentProteome oxidative carbonylation during oxidative stress-induced premature senescence of WI-38 human fibroblasts
2018, Mechanisms of Ageing and DevelopmentCitation Excerpt :This network includes such signalling proteins as NFκB, P38 MAPK, ERK, Akt, Pkc and AMPK among which P38 MAPK, ERK, Pkc and AMPK are also part of the network obtained for the 13 carbonylated proteins identified in SIPS WI-38 fibroblasts after a long recovery period of 30 days (Fig. 5C). Interestingly, all these signalling proteins have been previously implicated in the oxidative stress response and subsequent stress-induced premature senescence (Toussaint et al., 2002b; Bernard et al., 2004; Maruyama et al., 2009; Salminen et al., 2012; Boilan et al., 2013) which suggests that the “Oxi-proteome” might interfere with functional connections within these relevant signalling pathways. Interestingly, a decreased activation of the pro-survival Akt and ERK kinases has been previously reported for aged HDFs as compared to HDFs from young individuals upon exposure to hydrogen peroxide (Gurjala et al., 2005).
The impact of cellular senescence in skin ageing: A notion of mosaic and therapeutic strategies
2017, Biochemical PharmacologyCitation Excerpt :Finally, senescent cells use various methods of communication, which are discussed in Section 5. All these phenomena reflect the diversity of cellular senescence, leading to the mosaic of tissue ageing [36], which can be perceived in the skin (Fig. 2). Therefore, with age, the various skin cell types will enter into senescence.
Acrylamide induces accelerated endothelial aging in a human cell model
2015, Food and Chemical Toxicology