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Animals : an open access journal from MDPI2021; 11(7); doi: 10.3390/ani11071995

Characterization of an Ex Vivo Equine Endometrial Tissue Culture Model Using Next-Generation RNA-Sequencing Technology.

Abstract: Persistent mating-induced endometritis is a major cause of poor fertility rates in the mare. Endometritis can be investigated using an ex vivo equine endometrial explant system which measures uterine inflammation using prostaglandin F2α as a biomarker. However, this model has yet to undergo a wide-ranging assessment through transcriptomics. In this study, we assessed the transcriptomes of cultured endometrial explants and the optimal temporal window for their use. Endometrium harvested immediately post-mortem from native pony mares (n = 8) were sampled (0 h) and tissue explants were cultured for 24, 48 and 72 h. Tissues were stored in RNALater, total RNA was extracted and sequenced. Differentially expressed genes (DEGs) were defined using DESeq2 (R/Bioconductor). Principal component analysis indicated that the greatest changes in expression occurred in the first 24 h of culture when compared to autologous biopsies at 0 h. Fewer DEGs were seen between 24 and 48 h of culture suggesting the system was more stable than during the first 24 h. No genes were differentially expressed between 48 and 72 h but the low number of background gene expression suggested that explant viability was compromised after 48 h. ESR1, MMP9, PTGS2, PMAIP1, TNF, GADD45B and SELE genes were used as biomarkers of endometrial function, cell death and inflammation across tissue culture timepoints. STRING assessments of gene ontology suggested that DEGs between 24 and 48 h were linked to inflammation, immune system, cellular processes, environmental information processing and signal transduction, with an upregulation of most biomarker genes at 24 h. Taken together our observations indicated that 24-48 h is the optimal temporal window when the explant model can be used, as explants restore microcirculation, perform wound healing and tackle inflammation during this period. This key observation will facilitate the appropriate use of this as a model for further research into the equine endometrium and potentially the progression of mating-induced endometritis to persistent inflammation between 24 and 48 h.
Publication Date: 2021-07-03 PubMed ID: 34359123PubMed Central: PMC8300099DOI: 10.3390/ani11071995Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The authors of this research sought to better understand the optimal use of an ex vivo equine endometrial tissue culture model for investigating endometritis, a frequently observed fertility issue in mares. The findings suggest that the most effective window for using this model occurs between 24 and 48 hours, as the explants conduct essential functions such as microcirculation restoration, wound healing, and inflammation management during this period.

Study Objectives and Methodology

  • The main aim of the study was to evaluate the transcriptomes of cultured endometrial explants (tissue samples from the lining of the uterus) and to determine the best time window for their use in research.
  • The researchers obtained endometrium immediately post-mortem from eight pony mares and sampled them at different time intervals post-culture: 0 hours, 24 hours, 48 hours, and 72 hours.
  • The tissues were stored using RNALater, a preservation method for RNA, following which the total RNA was extracted and sequenced. They used a tool called DESeq2 to define differentially expressed genes (DEGs).

Key Findings

  • Principal Component Analysis, a statistical procedure used for simplifying complex data sets, showed that the biggest changes in expression occurred in the first 24 hours of culture, when compared with biopsies from the same tissue sample at 0 hours.
  • Fewer DEGs were observed between 24 and 48 hours of culture, suggesting greater stability of the system after the initial 24 hours.
  • No genes were differentially expressed between 48 and 72 hours. However, a notably low level of background gene expression suggested that explant viability might be compromised after 48 hours.
  • The researchers used a collection of specific genes as biomarkers to monitor endometrial function, cell death, and inflammation across the different timepoints.
  • The DEGs noted between 24 and 48 hours were associated with inflammation, immune system response, cellular processes, environmental information processing, and signal transduction, with most of these genes showing increased expression at 24 hours.

Implications and Conclusions

  • The findings indicate that the optimal temporal window for using the tissue culture model is between 24 and 48 hours, a period during which the explants restore microcirculation, aid wound healing and address inflammation.
  • The research, therefore, assists in identifying the most suitable use of this model for further study into equine endometrium and potential progression of mating-induced endometritis to persistent inflammation within the 24 to 48 hours timeframe.

Cite This Article

APA
Monteiro de Barros MR, Davies-Morel MCG, Mur LAJ, Creevey CJ, Alison RH, Nash DM. (2021). Characterization of an Ex Vivo Equine Endometrial Tissue Culture Model Using Next-Generation RNA-Sequencing Technology. Animals (Basel), 11(7). https://doi.org/10.3390/ani11071995

Publication

ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 11
Issue: 7

Researcher Affiliations

Monteiro de Barros, Maithê R
  • Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3FG, UK.
Davies-Morel, Mina C G
  • Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3FG, UK.
Mur, Luis A J
  • Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3FG, UK.
Creevey, Christopher J
  • Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast BT7 1NN, UK.
Alison, Roger H
  • Pathology Consultancy Services, Caerfyrddin Fach, Cilcennin, Lampeter SA48 8RN, UK.
Nash, Deborah M
  • Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3FG, UK.

Grant Funding

  • Ciência sem Fronteiras

Conflict of Interest Statement

All authors declare no conflict of interest. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, the writing of the manuscript, or in the decision to publish the results.

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