Establishment and characterization of bovine oviductal epithelial cells in culture and study of sperm binding capacity: "Feasibility studies towards the development of a semen fertility assessment assay for Norwegian red"
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The mammalian oviduct is the physiological site for key events in reproduction, such as capacitation of spermatozoa, fertilization and early embryonic developments. During passage through the oviduct, a fertilizing spermatozoon has to bind to and interact with epithelial cells at the caudal isthmus during the formation of functional sperm reservoir. Binding to these cells is thought to increase the fertile life span of sperm cells. In this study, bovine oviduct epithelial cells (BOECs) from NRF at estrus were cultured in monolayers and used to study sperm cells oviduct binding in vitro. The cultured cells were characterized by immunostaining and Real Time PCR was used to study the expression pattern of OVGP1 in cultured cells in the presence and absence of human chorionic gonadotrophin. Chlortetracycline staining was employed to study the capacitation status of sperm cells bound to monolayers of epithelial cells and Ca 2+ ionophore was used to induce sperm cell capacitation. Main findings demonstrate that (1) Primary BOECs cultures are a mixed population of cells; (2) Cultured BOECs loss the expression of OVGP1 over time during in vitro culture; (3) BOECs monolayers selectively bind uncapacitated sperm cells. These findings support the possible establishment of a sperm quality assessment assay through binding of sperm cells to BOECs monolayers.
Mastergradsoppgave i næringsrettet bioteknologi, Avdeling for lærerutdanning og naturvitenskap, Høgskolen i Hedmark, 2011. Master of applied and commercial biotechnology