The Long-Term Culture of Human Fibroblasts Reveals a Spectroscopic Signature of Senescence


Aging is a complex process which leads to progressive loss of fitness/capability/ability, increasing susceptibility to disease and, ultimately, death. Regardless of the organism, there are some features common to aging, namely, the loss of proteostasis and cell senescence. Mammalian cell lines have been used as models to study the aging process, in particular, cell senescence. Thus, the aim of this study was to characterize the senescence-associated metabolic profile of a long-term culture of human fibroblasts using Fourier Transform Infrared and Nuclear Magnetic Resonance spectroscopy. We sub-cultivated fibroblasts from a newborn donor from passage 4 to passage 17 and the results showed deep changes in the spectroscopic profile of cells over time. Late passage cells were characterized by a decrease in the length of fatty acid chains, triglycerides and cholesterol and an increase in lipid unsaturation. We also found an increase in the content of intermolecular beta-sheets, possibly indicating an increase in protein aggregation levels in cells of later passages. Metabolic profiling by NMR showed increased levels of extracellular lactate, phosphocholine and glycine in cells at later passages. This study suggests that spectroscopy approaches can be successfully used to study changes concomitant with cell senescence and validate the use of human fibroblasts as a model to monitor the aging process.



subject category

Biochemistry & Molecular Biology; Chemistry


Magalhaes, S; Almeida, I; Pereira, CD; Rebelo, S; Goodfellow, BJ; Nunes, A

our authors


This work was supported by Fundacao para a Ciencia e Tecnologia (FCT) I.P. (PIDDAC), European Regional Development Fund (FEDER) (project numbers UIDB/04501/2020; UIDB/50011/2020; UIDP/50011/2020; CENTRO-01-0145-FEDER-000003 and SFRH/BD/131820/2017 to SM). Image acquisition was performed in the LiMfacility of iBiMED, a node of PPBI (Portuguese Platform of BioImaging): POCI-01-0145-FEDER-022122.

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