Related Experiment Video
Updated: Jun 6, 2026
Quantitative and Qualitative Method for Sphingomyelin by LC-MS Using Two Stable Isotopically Labeled Sphingomyelin Species
Published on: May 7, 2018
Characterizing the effects of MSC lipid remodeling via sphingomyelinase licensing through morphological, metabolic,
S'Dravious A DeVeaux1, Daniel C Shah1, Kejie Rui2
1The Wallace H. Coulter Department of Biomedical Engineering, Georgia Tech and Emory, Atlanta, Georgia, USA; Petit Institute of Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia, USA.
Abstract:
Mesenchymal stromal cells (MSCs) are widely studied for their immunomodulatory and tissue reparative capabilities. However, clinical translation has been hampered by inconsistent efficacy and limited standardization in manufacturing. While cytokine-based priming methods, such as interferon-gamma (IFN-γ) stimulation, have shown promise in enhancing MSC potency, alternative approaches targeting distinct biological metabolism integral to secretome and membrane architecture have not been explored in MSCs. In this study, we investigate sphingomyelinase (SMase), an enzyme that generates ceramide from sphingomyelin, as a novel lipid-based priming strategy to modulate MSC response. Here, human MSCs were treated with SMase and high-content imaging. Morphologic profiling revealed that SMase-treated cells adopted a phenotype overlapping with IFN-γ-licensed MSCs, including increased cell compactness and solidity. Lipidomic analysis showed broad alterations in sphingolipid species, and dynamic flux estimation (DFE) modeling predicted distinct metabolic shifts in SMase-treated cells compared with untreated controls. These changes were sustained up to 38 hours post-stimulation, indicating stable metabolic reprogramming. Intracellular metabolic screening identified dynamic shifts in metabolite features leading to distinct profiles following 24-hour SMase treatment. SMase priming also altered the MSC secretome, enriching factors implicated in immune regulation. Functionally, SMase-primed MSCs retained the ability to suppress T-cell activation and promote anti-inflammatory macrophage phenotypes. Collectively, these findings demonstrate that SMase stimulation induces a durable, immunomodulatory-like state in MSCs through coordinated changes in lipid metabolism and secretory activity. This lipid-centric priming approach represents a promising alternative to cytokine-based licensing strategies and may support therapeutic MSC products.
