Impact of Pro-Inflammatory Cytokine Preconditioning on Metabolism and Extracellular Vesicles in Feline Mesenchymal Stromal Cells: A Preliminary Study

Maria Soltero-Rivera; Boaz Arzi; Lynda Bourebaba; Krzysztof Marycz

doi:10.1093/stmcls/sxaf014

Impact of Pro-Inflammatory Cytokine Preconditioning on Metabolism and Extracellular Vesicles in Feline Mesenchymal Stromal Cells: A Preliminary Study

Stem Cells. 2025 Apr 1:sxaf014. doi: 10.1093/stmcls/sxaf014. Online ahead of print.

Authors

Maria Soltero-Rivera^{1

2}, Boaz Arzi^{1

2}, Lynda Bourebaba³, Krzysztof Marycz^{1

2

3}

Affiliations

¹ Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California-Davis, Davis, CA, United States.
² Veterinary Institute for Regenerative Cures, School of Veterinary Medicine, University of California-Davis, Davis, CA, United States.
³ Department of Experimental Biology, Faculty of Biology and Animal Science, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.

PMID: 40168102
DOI: 10.1093/stmcls/sxaf014

Abstract

Background: Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have shown promise in treating inflammation. This study investigates whether preconditioning feline adipose-derived stem cells (FeASCs) with inflammatory cytokines, specifically IFN-γ and TNF-α, enhances the anti-inflammatory efficacy of MSC-derived EVs.

Objective: We hypothesize that cytokine-primed FeASCs will produce EVs with improved anti-inflammatory properties and that this preconditioning will affect mitochondrial dynamics to enhance EV therapy effectiveness.

Methods: FeASCs were exposed to TNF-α / IFN-γ combination to mimic a pro-inflammatory milieu favoring ASCs immunosuppressive phenotype. We analyzed morphological, metabolic, and immunomodulatory characteristics of native and cytokine-primed FeASCs. EVs were assessed for anti-inflammatory and mitochondrial-related markers. We also evaluated mitochondrial function and apoptosis markers in cytokine-primed cells.

Results: Cytokine priming led to significant morphological changes in FeASCs, including enhanced cell projections and increased apoptosis. EVs from cytokine-primed FeASCs exhibited a heightened immunomodulatory profile, with increased expression of both pro-inflammatory and anti-inflammatory mediators. Transcriptomic analysis of these EVs revealed the upregulation of genes associated with cell proliferation, survival, and apoptosis. Mitochondrial function was impaired in cytokine-primed cells, but mitochondrial morphology remained unchanged. EVs from these cells contained higher levels of mitochondrial-related transcripts, indicating a compensatory response.

Conclusion: Cytokine-primed FeASCs generate EVs with enhanced immunomodulatory potential, highlighting their therapeutic promise. However, further research is needed to validate their efficacy and safety and refine preconditioning strategies to optimize EV-based therapies for inflammatory conditions. These advancements could pave the way for broader applications in regenerative medicine.

Keywords: adipose; extracellular vesicle; feline; placental; regenerative medicine; stromal cell.