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

• Published in: International journal of molecular sciences Vol. 23; no. 10; p. 5830
• Main Authors: Magalhães, Sandra, Almeida, Idália, Pereira, Cátia D, Rebelo, Sandra, Goodfellow, Brian J, Nunes, Alexandra
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 23-05-2022; MDPI
• Subjects: Aging; Cell culture; Cell cycle; Cell growth; Cell lines; cell senescence; Cholesterol; Culture; Fibroblasts; Fourier transforms; FTIR spectroscopy; Glycine; human fibroblasts; Lactic acid; Lipids; Magnetic resonance spectroscopy; Metabolism; Metabolites; NMR; NMR spectroscopy; Nuclear magnetic resonance; Organisms; Phosphocholine; Protein interaction; Proteins; Senescence; Spectrum analysis; Triglycerides; aging; NMR spectroscopy; human fibroblasts; FTIR spectroscopy; cell senescence
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23105830

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 β-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.