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Senescence-Induced Lipidome Alterations in Mesenchymal Stromal Cells
Molly E Ogle1,2, Joseph L Corstvet3, Reesha K Vayalakkara1
1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 313 Ferst Drive, Atlanta, Georgia 30332, United States.
Abstract:
Mesenchymal stromal cells (MSCs) show great promise as a clinical treatment for a variety of diseases, but their susceptibility to senescence during culture reduces the therapeutic potential and limits cell expansion. In this study, we explored how MSC lipid metabolism is altered in culture over time using ultrahigh-performance liquid chromatography mass spectrometry. The proportion of cells with senescence-associated β-galactosidase (SA-β-gal) activity was evaluated during 12 days of culture expansion of MSCs from two human donors. Lipid profiles were evaluated in parallel using exact mass and tandem mass spectrometry spectral database matching to generate 237 unique lipid annotations. Lipid abundance generally increased across most lipid classes over serial culture; however, many changes were heterogeneous between donors. Despite donor differences, 12 lipids, including 4 triglycerides (TG), provided discrimination between cultures with less than 10% SA-β-gal+, those with 10-20% SA-β-gal+, and greater than 20% SA-β-gal+ senescence proportion regardless of donor. More specifically, TG composed of long-chain, highly unsaturated fatty acids was strongly associated with higher MSC senescence. These changes in bulk lipid profiles may inform future strategies to monitor early culture senescence during the expansion of MSCs.
Insights
Mesenchymal stromal cells (MSCs) accumulate specific lipid changes, particularly triglycerides, as they age in culture. These lipid alterations can help monitor and potentially mitigate senescence during MSC expansion for clinical use.
Area of Science:
- Cell Biology
- Biochemistry
- Metabolomics
Background:
- Mesenchymal stromal cells (MSCs) are promising for clinical applications.
- Cellular senescence limits MSC therapeutic potential and expansion.
- Understanding MSC culture dynamics is crucial for effective cell therapy.
Purpose of the Study:
- To investigate alterations in MSC lipid metabolism during culture expansion.
- To correlate lipid profiles with senescence markers in cultured MSCs.
- To identify potential lipid biomarkers for early senescence detection.
Main Methods:
- Cultured human MSCs from two donors for 12 days.
- Assessed senescence using senescence-associated β-galactosidase (SA-β-gal) activity.
- Analyzed lipid profiles using ultrahigh-performance liquid chromatography mass spectrometry (UHPLC-MS/MS).
- Identified 237 unique lipid species through spectral database matching.
Main Results:
- Lipid abundance generally increased across most classes during serial culture.
- Significant donor-to-donor heterogeneity was observed in lipid profiles.
- Twelve lipids, including four triglycerides (TGs), distinguished senescence levels (<10%, 10-20%, >20% SA-β-gal+).
- TGs with long-chain, unsaturated fatty acids correlated strongly with increased MSC senescence.
Conclusions:
- MSC lipid metabolism changes significantly during culture expansion.
- Specific lipid signatures, especially certain triglycerides, can indicate senescence.
- Lipid profiling offers a potential method for monitoring MSC culture quality and senescence.
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