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Updated: May 20, 2026

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
Published on: May 13, 2022
Protocol for isolation and characterization of small extracellular vesicles from human menstrual fluid using
Yiqun Tang1, Ruihan Zhou2, Caroline Frisendahl1
1WHO Collaborating Centre, Division for Neonatology, Obstetrics, Gynecology and Reproductive Health, Department of Women's and Children's Health, Karolinska Institutet, Biomedium A4, Solnavägen 9, 171 65 Stockholm, Sweden; Division of Gynecology and Reproductive Medicine, Karolinska University Hospital, Stockholm, Sweden.
Abstract:
Human menstrual fluid (MF) is rich in extracellular vesicles originating from peripheral blood components and local endometrial and immune cells. Here, we present a protocol to obtain MF-derived small extracellular vesicles (MF-sEVs) using dilution, low-speed centrifugation, filtration, and size-exclusion chromatography. We describe steps for characterizing pooled fractions by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and western blotting. This workflow yields high-quality MF-sEVs suitable for reproducible omics and functional assays, enabling robust biomarker discovery and therapeutic exploration in uterine disorders.

