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Updated: Dec 28, 2025
Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization
Published on: March 2, 2017
Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for
Paul Kelly1,2, Anne Barry-Reidy1,2, Amy Brewer1,2
1Comparative Immunology Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland.
Insights
This study details a reliable method for isolating and culturing primary bovine endometrial epithelial and stromal cells. These cells provide a relevant model for studying uterine immune responses and inflammation.
Area of Science:
- Reproductive Biology
- Immunology
- Cell Biology
Background:
- Reliable in-vitro models are crucial for studying the uterine immune response.
- Current protocols for isolating endometrial cells lack detail, leading to inconsistent results.
Purpose of the Study:
- To provide a detailed methodology for isolating and culturing distinct primary bovine endometrial epithelial and stromal cells.
- To demonstrate the utility of these cells in a physiologically relevant model for studying endometrial inflammation.
Main Methods:
- Bovine endometrial tissue was collected and separated using differential size filtering.
- Cell purity was confirmed via morphology, cytokeratin, and vimentin staining.
- Immune cell contamination was assessed using PCR for CD45.
Main Results:
- Isolation yielded significant numbers of epithelial cells and stromal fibroblasts.
- Both cell types exhibited increased inflammatory mediator expression (IL8, IL6, S100A8, S100A9) upon LPS challenge.
- Distinct inflammatory expression profiles were observed between epithelial and stromal cells.
Conclusions:
- A detailed and reproducible method for culturing primary bovine endometrial epithelial and stromal cells is presented.
- These primary cells serve as a valuable, physiologically relevant model for investigating endometrial inflammation and immune responses.
Objectives:
Isolation and culture of distinct primary endometrial cells are key to reliable in-vitro models to investigate the uterine immune response and optimse new disease interventions. Details on the isolation method and purity of distinct cell populations is lacking in currently available protocols leading to inconsistent results across laboratories.
Methods:
Bovine endometrial tissue from non-pregnant bovine uteri were collected immediately post-mortem and separated using differential size filtering. Isolations (n = 15) yielded an average of 3.1 × 105 ± 0.7 × 105 epithelial cells and 1.88 × 106 ± 5.44 × 105 stromal fibroblasts per uterine horn. Following expansion in culture, the purity of cell populations was confirmed using morphology and positive staining for cytokeratin and vimentin which identifies epithelial and stromal fibroblast populations, respectively. Using PCR, cDNA from both cell populations was negative for CD45, a marker of immune cells.
Results:
On challenge with a bacterial PAMP (LPS), epithelial and stromal fibroblasts showed a marked increase in the expression of the inflammatory mediators IL8, IL6, S100A8 and S100A9, with both cell populations displaying distinct expression profiles. Here we provide a detailed methodology on the culture of primary bovine endometrial epithelial and stromal cells and demonstrate these cells provide a physiologically relevant model for studies of endometrial inflammation and its regulation.
