title: Improved bovine embryo production in an oviduct-on-a-chip system: prevention of poly-spermic fertilization and parthenogenic activation.
abstract: The oviduct provides the natural micro-environment for gamete interaction, fertilization and early embryo development in mammals, such as the cow. In conventional culture systems, bovine oviduct epithelial cells (BOEC) undergo a rapid loss of essential differentiated cell properties; we aimed to develop a more physiological in vitro oviduct culture system capable of supporting fertilization. U-shaped chambers were produced using stereo-lithography and mounted with polycarbonate membranes, which were used as culture inserts for primary BOECs. Cells were grown to confluence and cultured at an air-liquid interface for 4 to 6 weeks and subsequently either fixed for immune staining, incubated with sperm cells for live-cell imaging, or used in an oocyte penetration study. Confluent BOEC cultures maintained polarization and differentiation status for at least 6 weeks. When sperm and oocytes were introduced into the system, the BOECs supported oocyte penetration in the absence of artificial sperm capacitation factors while also preventing polyspermy and parthenogenic activation, both of which occur in classical in vitro fertilization systems. Moreover, this "oviduct-on-a-chip" allowed live imaging of sperm-oviduct epithelium binding and release. Taken together, we describe for the first time the use of 3D-printing as a step further on bio-mimicking the oviduct, with polarized and differentiated BOECs in a tubular shape that can be perfused or manipulated, which is suitable for live imaging and supports in vitro fertilization.
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. b.m.gadella@uu.nl.
authors.forename: Marcia A M M
authors.initials: MAMM
authors.lastname: Ferraz
authors.affiliation: Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Heiko H W
authors.initials: HHW
authors.lastname: Henning
authors.affiliation: Department of Orthopedics, Utrecht Medical Center, Utrecht, The Netherlands and Utrecht Biofabrication Facility, Utrecht Medical Center, Utrecht, The Netherlands.
authors.forename: Pedro F
authors.initials: PF
authors.lastname: Costa
authors.affiliation: Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands and Department of Orthopedics, Utrecht Medical Center, Utrecht, The Netherlands and Utrecht Biofabrication Facility, Utrecht Medical Center, Utrecht, The Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: Malda
authors.affiliation: Department of Orthopedics, Utrecht Medical Center, Utrecht, The Netherlands and Utrecht Biofabrication Facility, Utrecht Medical Center, Utrecht, The Netherlands.
authors.forename: Ferry P
authors.initials: FP
authors.lastname: Melchels
authors.affiliation: Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Wubbolts
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. b.m.gadella@uu.nl and Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Tom A E
authors.initials: TAE
authors.lastname: Stout
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. b.m.gadella@uu.nl.
authors.forename: Peter L A M
authors.initials: PLAM
authors.lastname: Vos
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. b.m.gadella@uu.nl and Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Bart M
authors.identifier: 0000-0001-9618-1261
authors.initials: BM
authors.lastname: Gadella
mesh_terms: D000818:Animals; D002417:Cattle; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D004867:Equipment Design; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005307:Fertilization in Vitro / Q000662:veterinary*; D056656:Lab-On-A-Chip Devices / Q000662:veterinary*; D008297:Male; D018613:Microscopy, Confocal; D010057:Oviducts / Q000166:cytology*; D010312:Parthenogenesis / Q000502:physiology*; D066330:Printing, Three-Dimensional; D013094:Spermatozoa / Q000166:cytology*