DOI: 10.1177/22840265261458937 ISSN: 2284-0265

Spectral flow cytometry of plasma and cervicovaginal fluid extracellular vesicles for diagnostic endometriosis biomarkers

E. S. J. Paterson, S. Scheck, S. McDowell, G. Roopi, N. Bedford, J. E. Girling, C. E. Henry
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Introduction:

Evidence for the role of extracellular vesicles (EVs) in endometriosis pathophysiology suggests EVs may be a novel avenue for biomarker exploration. This study employed small particle flow cytometry to investigate plasma EVs with endometrial stromal cell markers and cervicovaginal fluid EVs with immunological receptors as diagnostic biomarkers of endometriosis.

Methods:

Blood and cervicovaginal fluid were collected from consenting patients undergoing surgery for suspected endometriosis. EVs were isolated from plasma ( n  = 15 per group) and cervicovaginal fluid ( n  = 20 per group), stained overnight with antibody probes, labelled with Aco-430 membrane stain then analysed by flow cytometry (Cytek Aurora). The proportions and antigen density of positive EVs were compared between people with endometriosis and symptomatic controls.

Results:

Measuring triple-positive EVs was deemed unsuitable given the variability in staining when multiplexing antibodies. Small proportions of antigen positive EVs were detected for all proteins investigated, however, no differences were observed for CD10 + , CD90 + , CD10 + CD90 + and CD140b + plasma EVs, or CCR2 + , CD55 + , CD59 + and CD82 + cervicovaginal fluid EVs.

Conclusions:

Although the EV populations did not distinguish endometriosis from symptomatic controls, this study provides valuable methodological insights into EV phenotyping to support future biomarker research and advance understanding of EV-mediated mechanisms in endometriosis.

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