Investigation of MSC potency metrics via integration of imaging modalities with lipidomic characterization
Priyanka Priyadarshani,
Alexandria Van Grouw,
Adrian Ross Liversage,
Kejie Rui,
Arina Nikitina,
Kayvan Forouhesh Tehrani,
Bhavay Aggarwal,
Steven L. Stice,
Saurabh Sinha,
Melissa L. Kemp,
Facundo M. Fernández,
Luke J. Mortensen
Affiliations
Priyanka Priyadarshani
School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA; Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA
Alexandria Van Grouw
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA
Adrian Ross Liversage
School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA; Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA
Kejie Rui
School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA; Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA
Arina Nikitina
School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA 30332, USA
Kayvan Forouhesh Tehrani
Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Champaign, IL 61820, USA
Bhavay Aggarwal
The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University, Atlanta, GA 30332, USA
Steven L. Stice
Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA
Saurabh Sinha
The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University, Atlanta, GA 30332, USA
Melissa L. Kemp
The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University, Atlanta, GA 30332, USA
Facundo M. Fernández
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA
Luke J. Mortensen
School of Chemical, Materials, and Biomedical Engineering, University of Georgia, Athens, GA 30602, USA; Regenerative Bioscience Center, Rhodes Center for ADS, University of Georgia, Athens, GA 30602, USA; Corresponding author
Summary: 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.
