Faculty, Staff and Student Publications

Language

English

Publication Date

1-1-2026

Journal

Journal of the American College of Surgeons

DOI

10.1097/XCS.0000000000001591

PMID

41051108

PMCID

PMC12704670

PubMedCentral® Posted Date

10-6-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Background: Mesenchymal stromal cell (MSC)-derived extracellular vesicles (EVs) offer a promising acellular therapy for immune modulation, but clinical translation is hindered by variability and storage constraints. Lyophilization (freeze-drying) could enable shelf-stable EV therapeutics, although its effects on biological activity remain incompletely defined.

Study design: We compared the immunomodulatory effects of frozen and lyophilized EVs from bone marrow (BM), adipose tissue (AD), and umbilical cord (UC) MSCs using lipopolysaccharide-stimulated rodent splenocytes and microglial models and human peripheral blood mononuclear cells (PBMCs). Variables included batch scale (small vs large), cytokine priming, and MSC source. Tumor necrosis factor alpha (TNF-α) secretion was measured as a marker of inflammation.

Results: Lyophilized small-batch BM-derived MSC EVs consistently suppressed TNF-α in rodent splenocytes and microglia. Large-batch and cytokine-stimulated EVs showed more variable responses. UC- and AD-derived EVs elicited inconsistent effects in rodent splenocytes. In contrast, lyophilized UC- and AD-derived EVs on human PBMCs exhibited reproducible suppression of TNF-α across 3 donors.

Conclusions: Lyophilized MSC-EVs-particularly from UC MSCs-retain immunomodulatory function and perform comparably or better than frozen counterparts. Although rodent models revealed significant variability, human PBMCs demonstrated consistent cytokine suppression. These findings support the feasibility of lyophilized EVs as scalable, shelf-stable therapeutics for inflammatory and neuroinflammatory conditions and underscore the need for standardized manufacturing and potency assays.

Keywords

Extracellular Vesicles, Freeze Drying, Animals, Mesenchymal Stem Cells, Humans, Adipose Tissue, Leukocytes, Mononuclear, Tumor Necrosis Factor-alpha, Rats, Neuroinflammatory Diseases, Mice, Umbilical Cord, Spleen, Microglia

Published Open-Access

yes

Download

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.