Characterization of an Ex Vivo Equine Endometrial Tissue Culture Model Using Next-Generation RNA-Sequencing Technology.
Authors: Monteiro de Barros Maithê R, Davies-Morel Mina C G, Mur Luis A J, Creevey Christopher J, Alison Roger H, Nash Deborah M
Journal: Animals : an open access journal from MDPI
Summary
# Editorial Summary Persistent mating-induced endometritis remains a significant fertility challenge in mares, and ex vivo endometrial tissue culture offers a valuable research tool for understanding inflammatory responses; however, the temporal stability and gene expression characteristics of this model required comprehensive evaluation. Researchers cultured endometrial explants from eight pony mares at four timepoints (0, 24, 48 and 72 hours post-harvest) and performed RNA-sequencing to characterise differential gene expression, identifying key inflammatory biomarkers including PTGS2 (prostaglandin synthase), TNF and SELE across the culture period. The 24–48 hour window emerged as optimal for experimental work, with greatest transcriptional changes occurring in the first 24 hours as tissues underwent microcirculation restoration and wound healing, whilst explant viability declined significantly beyond 48 hours despite minimal gene expression changes between 48–72 hours. Gene ontology analysis confirmed that DEGs between 24–48 hours clustered around inflammation, immune signalling and cellular processes, with most biomarker genes showing upregulation at 24 hours. For practitioners utilising this model in endometritis research, these findings establish precise temporal boundaries for experimental design and validate the system's capacity to investigate how acute post-mating inflammation progresses to persistent endometritis, ultimately refining our understanding of subfertility mechanisms in mares.
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Practical Takeaways
- •This ex vivo model provides a validated research tool for investigating equine endometritis mechanisms; clinicians should be aware of its 24-48 h optimal window when interpreting related published findings
- •The identified biomarkers and temporal dynamics offer potential for developing diagnostic or prognostic tools to assess endometrial inflammation and predict progression to persistent mating-induced endometritis
- •Understanding the natural inflammatory and wound-healing response in the first 48 h of culture may inform timing of therapeutic interventions in mares with documented endometritis
Key Findings
- •Greatest transcriptomic changes occurred in the first 24 h of endometrial explant culture compared to 0 h controls
- •No differential gene expression detected between 48 and 72 h, indicating compromised explant viability after 48 h
- •The 24-48 h window represents the optimal temporal window for explant use as tissue undergoes microcirculation restoration, wound healing, and inflammation response
- •Biomarker genes (ESR1, MMP9, PTGS2, PMAIP1, TNF, GADD45B, SELE) show upregulation at 24 h, with gene ontology linked to inflammation, immune function, and signal transduction