title: Development of an in vitro oviduct epithelial explants model for studying sperm-oviduct binding in the buffalo.
abstract: In this study, we developed an in vitro model for studying sperm-oviduct binding in the buffalo. Oviduct explants were prepared by overnight culture of epithelial cells in TCM-199 medium under 5% CO2at 38.5 °C. Cryopreserved spermatozoa from buffalo bulls (n = 4) were incubated with the oviduct explants, and the sperm-oviduct explants complex was stained with JC-1. The effect of sperm concentration (2, 3 and 4 million), size of the oviduct explants (<0.2, 0.2-0.3, 0.3-0.4 and >0.4 mm2) and time of incubation (1 hr and 4 hr) on binding index (BI-number of sperm bound to unit area of explants) was studied. No significant difference was observed in the BI among <0.2, 0.2-0.3 and 0.3-0.4 mm2size of explants; however, the BI decreased significantly (p < .05) when the size of explants exceeded 0.4 mm2. The BI decreased significantly (p < .05) when the sperm concentration was increased to 4 million, while the duration of incubation did not have any significant effect on the BI. The interaction of bulls with explants size, sperm concentration and incubation time was not significant. The developed assay has the potential to be used as an in vitro model for studying sperm-oviduct binding in the buffalo.
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: K K
authors.initials: KK
authors.lastname: Saraf
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: A
authors.identifier: http://orcid.org/0000-0002-8061-3477
authors.initials: A
authors.lastname: Kumaresan
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: S
authors.initials: S
authors.lastname: Nayak
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: S
authors.initials: S
authors.lastname: Chhillar
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: L
authors.initials: L
authors.lastname: Sreela
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: S
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Theriogenology Lab, Animal Reproduction, Gynaecology & Obstetrics, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: U K
authors.initials: UK
authors.lastname: Tripathi
authors.affiliation: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: T K
authors.initials: TK
authors.lastname: Datta
authors.affiliation: Artificial Breeding Research Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors.forename: T K
authors.initials: TK
authors.lastname: Mohanty
mesh_terms: D000818:Animals; D002020:Buffaloes; D018929:Cell Culture Techniques; D015925:Cryopreservation; D004847:Epithelial Cells / Q000502:physiology; D005260:Female; D005298:Fertility; D008297:Male; D010057:Oviducts / Q000502:physiology*; D012662:Semen Preservation; D013094:Spermatozoa / Q000502:physiology*
keywords: JC-1; buffalo; oviduct explants; sperm binding