Investigation of MSC potency metrics via integration of imaging modalities with lipidomic characterization

• Published in: Cell reports (Cambridge) 2024-08, Vol.43 (8), p.114579, Article 114579
• Main Authors: Priyadarshani, Priyanka, Van Grouw, Alexandria, Liversage, Adrian Ross, Rui, Kejie, Nikitina, Arina, Tehrani, Kayvan Forouhesh, Aggarwal, Bhavay, Stice, Steven L., Sinha, Saurabh, Kemp, Melissa L., Fernández, Facundo M., Mortensen, Luke J.
• Format: Article
• Language: English
• Subjects: Ceramides - metabolism; CP: Immunology; CP: Metabolism; Humans; Interferon-gamma - metabolism; Lipid Metabolism; Lipidomics - methods; Lipids - analysis; Lipids - chemistry; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; Single-Cell Analysis - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2024.114579

Mesenchymal stem/stromal cell (MSC) therapies have had limited success so far in clinical trials due in part to heterogeneity in immune-responsive phenotypes. Therefore, techniques to characterize these properties of MSCs are needed during biomanufacturing. Imaging cell shape, or morphology, has been found to be associated with MSC immune responsivity—but a direct relationship between single-cell morphology and function has not been established. We used label-free differential phase contrast imaging and matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to evaluate single-cell morphology and explore relationships with lipid metabolic immune response. In interferon gamma (IFN-γ)-stimulated MSCs, we found higher lipid abundances from the ceramide-1-phosphate (C1P), phosphatidylcholine (PC), LysoPC, and triglyceride (TAG) families that are involved in cell immune function. Furthermore, we identified differences in lipid signatures in morphologically defined MSC subpopulations. The use of single-cell optical imaging coupled with single-cell spatial lipidomics could assist in optimizing the MSC production process and improve mechanistic understanding of manufacturing process effects on MSC immune activity and heterogeneity. [Display omitted] •Identified single-cell lipid signatures of morphologically distinct MSC subpopulations•Specific morphology features and lipids are associated in immune activated cells•Key immune-responsive lipid pathways alter morphology and lead to a decline in function Priyadarshani et al. investigated the relationship between the morphological and lipidomic profiles of single-cell MSCs. Their study revealed that, by analyzing these profiles at the single-cell level, they could identify specific combinations of morphological and lipidomic characteristics in IFN-γ-stimulated MSCs that distinguish functional MSC subpopulations from non-functional ones.