Bovine oviductal monolayers cultured under three-dimension conditions secrete factors able to release spermatozoa adhering to the tubal reservoir in vitro

Different in vitro models have been developed to understand the interaction of gametes and embryos with the maternal reproductive tract. We recently showed that bovine oviductal monolayers three-dimensionally cultured in Gray's medium on collagen-coated microporous polycarbonate inserts under liquid-air interface conditions are well polarized, develop cilia, remain viable for at least 3 weeks postconfluence, and mantain the viability of bound spermatozoa significantly better than bidimensionally cultured monolayers. Herein, we used these culture conditions to understand whether: (1) spermatozoa adhering to three-dimensionally cultured oviductal monolayers can be released by heparin or penicillamine as previously shown with bidimensionally cultured oviductal monolayers and explants; and (2) media conditioned by three-dimensionally cultured oviductal monolayers were able to release spermatozoa adhering to oviductal explants. Findings demonstrated that (1) spermatozoa adhering to three-dimensionally cultured oviductal monolayers are readily released by heparin and penicillamine, (2) media conditioned by three-dimensionally cultured oviductal monolayers are able to release spermatozoa bound to oviductal explants, (3) do not depress sperm motility and viability, (4) they improve sperm kinetics, and (5) promote binding to the zona pellucida. In conclusion, in vitro data suggest that the release of spermatozoa adhering to the oviductal reservoir in vivo can be triggered by factors secreted by the oviduct itself that induce sperm capacitation.