Raman and infrared spectroscopy reveal that proliferating and quiescent human fibroblast cells age by biochemically similar but not identical processes

• Published in: PloS one 2018-12, Vol.13 (12), p.e0207380-e0207380
• Main Authors: Eberhardt, Katharina, Matthäus, Christian, Marthandan, Shiva, Diekmann, Stephan, Popp, Jürgen
• Format: Article
• Language: English
• Subjects: Age; Aging; Aging (artificial); Amino acids; Apoptosis; Autophagy; Biochemistry; Biology and Life Sciences; Cell cycle; Cell division; Cell growth; Cell Proliferation; Cellular Senescence; Composition; Cultivation; Deoxyribonucleic acid; DNA; DNA damage; Fibroblasts; Fibroblasts - cytology; Fibroblasts - metabolism; Gene expression; Homeostasis; Humans; Infrared spectroscopy; Kinases; Laboratories; Lipids; Lipmann, Fritz; Medicine and Health Sciences; Molecular biology; Multivariate statistical analysis; Nucleic acids; Photonics; Physical chemistry; Physical Sciences; Protein structure; Proteins; Research and Analysis Methods; Schiller, Johann Christoph Friedrich von (1759-1805); Secondary structure; Senescence; Skin; Spectrophotometry, Infrared; Spectroscopic analysis; Spectroscopy; Spectrum Analysis, Raman; Statistical analysis; Time Factors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0207380

Dermal fibroblast cells can adopt different cell states such as proliferation, quiescence, apoptosis or senescence, in order to ensure tissue homeostasis. Proliferating (dividing) cells pass through the phases of the cell cycle, while quiescent and senescent cells exist in a non-proliferating cell cycle-arrested state. However, the reversible quiescence state is in contrast to the irreversible senescence state. Long-term quiescent cells transit into senescence indicating that cells age also when not passing through the cell cycle. Here, by label-free in vitro vibrational spectroscopy, we studied the biomolecular composition of quiescent dermal fibroblast cells and compared them with those of proliferating and senescent cells. Spectra were examined by multivariate statistical analysis using a PLS-LDA classification model, revealing differences in the biomolecular composition between the cell states mainly associated with protein alterations (variations in the side chain residues of amino acids and protein secondary structure), but also within nucleic acids and lipids. We observed spectral changes in quiescent compared to proliferating cells, which increased with quiescence cultivation time. Raman and infrared spectroscopy, which yield complementary biochemical information, clearly distinguished contact-inhibited from serum-starved quiescent cells. Furthermore, the spectra displayed spectral differences between quiescent cells and proliferating cells, which had recovered from quiescence. This became more distinct with increasing quiescence cultivation time. When comparing proliferating, (in particular long-term) quiescent and senescent cells, we found that Raman as well as infrared spectroscopy can separate these three cellular states from each other due to differences in their biomolecular composition. Our spectroscopic analysis shows that proliferating and quiescent fibroblast cells age by similar but biochemically not identical processes. Despite their aging induced changes, over long time periods quiescent cells can return into the cell cycle. Finally however, the cell cycle arrest becomes irreversible indicating senescence.