Isolation, culture, and characterization of equine oviduct epithelial cells in vitro.
Abstract: Oviduct epithelial cells (OEC) increasingly are used to support embryonic development and to study gamete interactions with the female reproductive tract in vitro. This series of experiments was designed to characterize monolayers derived from oviduct epithelium. Epithelial cells harvested from the isthmus and ampulla of the oviducts of five estrous mares were cultured with or without the basal lamina extract, Matrigel. Within each group OEC were cultured in the presence of either estradiol-17 beta or a carrier control. All groups were subcultured three times. Epithelial cell morphology and function were examined by microscopy, analysis of secreted proteins, and immunocytochemistry. Epithelial cells attached more rapidly and reached confluence sooner when cultured on Matrigel than in uncoated wells. Cells showed variable evidence of ciliary activity up to 12 days in primary culture. Cells grown on Matrigel had a more polarized appearance in primary culture than those in uncoated wells, although no morphologic difference between anatomic site of origin or between steroid treated groups was noted. Anatomic site of origin had no effect, and steroid treatment had minimal effects, on patterns of secreted proteins. However, some differences were noted in protein secretion between cells grown with or without Matrigel. These data suggest that culture substrate may affect structure and function of OEC monolayers.
Publication Date: 1995-08-01 PubMed ID: 7576614DOI: 10.1002/mrd.1080410409Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study investigates the use of equine oviduct epithelial cells (OEC) in supporting embryonic development and explores their interactions with the female reproductive tract during in vitro studies. The researchers found that the cells attached more quickly and achieved confluence sooner on a particular culture substrate, Matrigel.
Objective and Methodology
- The objective of this research paper was to characterize the monolayers derived from oviduct epithelium and study how their culture substrate can affect their structure and function.
- The study used epithelial cells harvested from the isthmus and ampulla of the oviducts of five estrous mares. These cells were cultured with or without Matrigel, a basal lamina extract. Further, the researchers introduced either estradiol-17 beta (a type of estrogen) or a carrier control to each group of oviduct epithelial cells.
- Cell morphology and function were analyzed using microscopy, observation of secreted proteins, and immunocytochemistry.
Findings
- The researchers found that epithelial cells attached more quickly and reached confluence (a state of completeness or full development in cell cultures) sooner when they were cultured on Matrigel as compared to uncoated wells.
- These cells showed varying degrees of ciliary activity (motion of tiny, hair-like structures on the cell surface) until 12 days in primary culture. Cells grown on Matrigel appeared more polarized (organized in a specific direction) in primary culture than those cultured in uncoated wells.
- No morphological difference was noted between the epithelial cells according to their anatomical site of origin or whether they were treated with steroids.
- Pattern of proteins secreted by the cells were not notably affected by the anatomical site of origin or by the steroid treatment. However, some variance was observed in protein secretion between cells grown with or without Matrigel.
Conclusion
- The study concludes that the culture substrate (Matrigel or uncoated wells) may affect the structure and function of monolayers of OEC. The substrate therefore could potentially play a significant role in supporting embryonic development during in vitro studies.
Cite This Article
APA
Thomas PG, Ignotz GG, Ball BA, Miller PG, Brinsko SP, Currie B.
(1995).
Isolation, culture, and characterization of equine oviduct epithelial cells in vitro.
Mol Reprod Dev, 41(4), 468-478.
https://doi.org/10.1002/mrd.1080410409 Publication
Researcher Affiliations
- Department of Clinical Sciences, Cornell University, Ithaca, New York, USA.
MeSH Terms
- Animals
- Cell Adhesion
- Cell Polarity
- Cell Separation
- Cells, Cultured
- Cilia / ultrastructure
- Collagen
- Culture Techniques / methods
- Drug Combinations
- Epithelial Cells
- Epithelium / drug effects
- Estradiol / pharmacology
- Fallopian Tubes / cytology
- Female
- Horses / anatomy & histology
- Laminin
- Microscopy, Electron
- Proteoglycans
Citations
This article has been cited 4 times.- Leemans B, Bromfield EG, Stout TAE, Vos M, Van Der Ham H, Van Beek R, Van Soom A, Gadella BM, Henning H. Developing a reproducible protocol for culturing functional confluent monolayers of differentiated equine oviduct epithelial cells†. Biol Reprod 2022 Apr 26;106(4):710-729.
- Rajagopal M, Tollner TL, Finkbeiner WE, Cherr GN, Widdicombe JH. Differentiated structure and function of primary cultures of monkey oviductal epithelium. In Vitro Cell Dev Biol Anim 2006 Sep-Oct;42(8-9):248-54.
- Baghaban Eslami Nejad MR, Rezazadeh Valojerdi M, Kazemi Ashtiani S. A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development. J Exp Clin Assist Reprod 2005 Apr 19;2(1):7.
- Leemans B, Gadella BM, Marchand JHEAM, Van Soom A, Stout TAE. Induction of in vivo-like ciliation in confluent monolayers of re-differentiated equine oviduct epithelial cells†. Biol Reprod 2024 Sep 14;111(3):580-599.
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