Tags

Type your tag names separated by a space and hit enter

Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies.
Vet Res Commun. 2020 Feb; 44(1):29-39.VR

Abstract

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.

Authors+Show Affiliations

Comparative Immunology Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland. Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Ireland.Comparative Immunology Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland. Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Ireland.Comparative Immunology Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland. Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Ireland.Animal & Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Teagasc, Grange, Ireland. kieran.meade@teagasc.ie.Comparative Immunology Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland. School of Medicine, Trinity College Dublin, Dublin, Ireland.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32086740

Citation

Kelly, Paul, et al. "Improved Filtration Method to Isolate Pure Populations of Primary Bovine Endometrial Epithelial and Stromal Cells for Immunological Studies." Veterinary Research Communications, vol. 44, no. 1, 2020, pp. 29-39.
Kelly P, Barry-Reidy A, Brewer A, et al. Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies. Vet Res Commun. 2020;44(1):29-39.
Kelly, P., Barry-Reidy, A., Brewer, A., Meade, K. G., & O'Farrelly, C. (2020). Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies. Veterinary Research Communications, 44(1), 29-39. https://doi.org/10.1007/s11259-020-09770-3
Kelly P, et al. Improved Filtration Method to Isolate Pure Populations of Primary Bovine Endometrial Epithelial and Stromal Cells for Immunological Studies. Vet Res Commun. 2020;44(1):29-39. PubMed PMID: 32086740.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies. AU - Kelly,Paul, AU - Barry-Reidy,Anne, AU - Brewer,Amy, AU - Meade,Kieran G, AU - O'Farrelly,Cliona, Y1 - 2020/02/21/ PY - 2020/01/16/received PY - 2020/01/31/accepted PY - 2020/2/23/pubmed PY - 2020/2/23/medline PY - 2020/2/23/entrez KW - Epithelial cell KW - Innate immune response KW - Primary culture KW - Stromal fibroblast SP - 29 EP - 39 JF - Veterinary research communications JO - Vet. Res. Commun. VL - 44 IS - 1 N2 - 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. SN - 1573-7446 UR - https://www.unboundmedicine.com/medline/citation/32086740/Improved_filtration_method_to_isolate_pure_populations_of_primary_bovine_endometrial_epithelial_and_stromal_cells_for_immunological_studies L2 - https://doi.org/10.1007/s11259-020-09770-3 DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.