Equine oviduct explant culture: a basic model to decipher embryo–maternal communication
| Autor: | Karen Goossens, Katrien Smits, Katharina D'Herde, Hilde Nelis, Kevin Braeckmans, Katrien Forier, Lynn Vandenberghe, Bart Leemans, Ann Van Soom, Luc Peelman |
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| Rok vydání: | 2014 |
| Předmět: |
Time Factors
Stromal cell Cell Culture Techniques Gene Expression Apoptosis Reproductive technology Biology Models Biological Endocrinology Microscopy Electron Transmission In Situ Nick-End Labeling Genetics medicine Animals Horses Lactic Acid Molecular Biology Fallopian Tubes Lamina propria Mucous Membrane Reverse Transcriptase Polymerase Chain Reaction Epithelial Cells Embryo culture Embryo Embryo Mammalian Cell Hypoxia Epithelium Cell biology Glucose medicine.anatomical_structure Reproductive Medicine Immunology Oviduct Female Animal Science and Zoology Stromal Cells Transcriptome Developmental Biology Biotechnology Explant culture |
| Zdroj: | Reproduction, Fertility and Development. 26:954 |
| ISSN: | 1031-3613 |
| DOI: | 10.1071/rd13089 |
| Popis: | Equine embryos remain for 6 days in the oviduct and thus there is a need for an in vitro model to study embryo–oviductal interactions in the horse, since this subtle way of communication is very difficult to analyse in vivo. Until now, no equine oviduct explant culture model has been characterised both morphologically and functionally. Therefore, we established a culture system for equine oviduct explants that maintained epithelial morphology during 6 days of culture, as revealed by light microscopy and transmission electron microscopy. We demonstrated the presence of highly differentiated, tall columnar, pseudostratified epithelium with basal nuclei, numerous nucleoli, secretory granules and apical cilia, which is very similar to the in vivo situation. Both epithelium and stromal cells originating from the lamina propria are represented in the explants. Moreover, at least 98% of the cells remained membrane intact and fewer than 2% of the cells were apoptotic after 6 days of culture. Although dark-cell degeneration, which is a hypoxia-related type of cell death, was observed in the centre of the explants, quantitative real-time PCR failed to detect upregulation of the hypoxia-related marker genes HIF1A, VEGFA, uPA, GLUT1 and PAI1. Since the explants remained morphologically and functionally intact and since the system is easy to set up, it appears to be an excellent tool for proteome, transcriptome and miRNome analysis in order to unravel embryo–maternal interactions in the horse. |
| Databáze: | OpenAIRE |
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