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

Gualtieri, Roberto; Mollo, Valentina; Braun, Sabrina; Barbato, Vincenza; Fiorentino, Ilaria; Talevi, Riccardo

doi:10.1016/j.theriogenology.2012.10.014

Cited by 17 publications

(19 citation statements)

References 29 publications

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“…It is possible to ligate and excise the oviduct from experimental animals and given reproductive stages and to fix the tissues for histological or other microscopic investigation 13,40. It is also possible to harvest epithelial cells from recovered oviducts.…”

Section: Studying Oviduct Functionmentioning

confidence: 99%

“…4a). 2-D culture of OECs is hampered by a rapid loss of typical differentiated OEC properties, such as ciliation, columnar cell morphology, cell polarity, secretory granules and bulbous protrusions 8,39,40,47,48,101,104,115. The use of 2-D culture was nevertheless a useful first step in trying to understand the roles of the oviduct during gamete interaction and early embryo development.…”

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

“…4b). Oviduct explants are aggregates of epithelial and stromal cells that organize to form motile, everted vesicles with active cilia on the outer surface 40,66,86. OECs in oviduct explants are able to maintain their differentiated morphology as characterized by secondary cilia, numerous mitochondria and rough endoplasmic reticulum,86 and expression of oviductal epithelial cell markers such as oviductal glycoprotein 1 (OVGP1), glutathione peroxidase 4 (GPX4) and forkhead box protein 1 (FOXJ1) 71.…”

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

“…3c and 3d) have been developed in recent years. This technique allows cultured OECs to retain their polarized columnar epithelial cell characteristics, and has been applied successfully to OECs from various mammalian species 14,16,34,40,92–94. Within the inserts, seeded OECs first form a confluent layer on the porous membrane.…”

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

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Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

et al. 2016

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The oviduct was long considered a largely passive conduit for gametes and embryos. However, an increasing number of studies into oviduct physiology have demonstrated that it specifically and significantly influences gamete interaction, fertilization and early embryo development. While oviduct epithelial cell (OEC) function has been examined during maintenance in conventional tissue culture dishes, cells seeded into these two-dimensional (2-D) conditions suffer a rapid loss of differentiated OEC characteristics, such as ciliation and secretory activity. Recently, three-dimensional (3-D) cell culture systems have been developed that make use of cell inserts to create basolateral and apical medium compartments with a confluent epithelial cell layer at the interface. Using such 3-D culture systems, OECs can be triggered to redevelop typical differentiated cell properties and levels of tissue organization can be developed that are not possible in a 2-D culture. 3-D culture systems can be further refined using new micro-engineering techniques (including microfluidics and 3-D printing) which can be used to produce ‘organs-on-chips’, i.e. live 3-D cultures that bio-mimic the oviduct. In this review, concepts for designing bio-mimic 3-D oviduct cultures are presented. The increased possibilities and concomitant challenges when trying to more closely investigate oviduct physiology, gamete activation, fertilization and embryo production are discussed.

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Section: Studying Oviduct Functionmentioning

confidence: 99%

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

See 2 more Smart Citations

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

et al. 2016

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“…In those experiments, capacitation had been stimulated by heparin [8], which could have prevented attachment by interacting with the BSP proteins on the surface of sperm [9], because these proteins each contain two heparin-binding sites [10, 11]. Heparin has been shown to release sperm bound to cultured oviductal epithelium in vitro [12]. …”

Section: Introductionmentioning

confidence: 99%

Dynamics of Bovine Sperm Interaction with Epithelium Differ Between Oviductal Isthmus and Ampulla

Ardón

Markello

et al. 2016

Biology of Reproduction

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In mammals, many sperm that reach the oviduct are held in a reservoir by binding to epithelium. To leave the reservoir, sperm detach from the epithelium; however, they may bind and detach again as they ascend into the ampulla toward oocytes. In order to elucidate the nature of binding interactions along the oviduct, we compared the effects of bursts of strong fluid flow (as would be caused by oviductal contractions), heparin, and hyperactivation on detachment of bovine sperm bound in vitro to epithelium on intact folds of isthmic and ampullar mucosa. Intact folds of oviductal mucosa were used to represent the strong attachments of epithelial cells to each other and to underlying connective tissue that exist in vivo. Effects of heparin on binding were tested because heparin binds to the Binder of SPerm (BSP) proteins that attach sperm to oviductal epithelium. Sperm bound by their heads to beating cilia on both isthmic and ampullar epithelia and could not be detached by strong bursts of fluid flow. Addition of heparin immediately detached sperm from isthmic epithelium but not ampullar epithelium. Addition of 4-aminopyridine immediately stimulated hyperactivation of sperm but did not detach them from isthmic or ampullar epithelium unless added with heparin. These observations indicate that the nature of binding of sperm to ampullar epithelium differs from that of binding to isthmic epithelium; specifically, sperm bound to isthmic epithelium can be detached by heparin alone, while sperm bound to ampullar epithelium requires both heparin and hyperactivation to detach from the epithelium.

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Toll‐like receptor 2 mediates the immune response of the bovine oviductal ampulla to sperm binding

Morillo

Akthar

Fiorenza

et al. 2020

Molecular Reproduction Devel

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We previously reported that sperm binding to cultured bovine oviduct epithelial cells induces an anti-inflammatory immune response. Now we have developed a differentiated explant model to focus on the oviductal ampulla, where fertilization occurs, and to study the effect of sperm capacitation on the immune response. We used heparin to stimulate bovine sperm capacitation. Fluorescence imaging showed that 5,5ʹ,6,6ʹ-tetrachloro-1,1ʹ,3,3ʹ-tetraethylbenzimidazolyl-carbocyanine iodidelabeled sperm pretreated with (Hep (+)) or without (Hep (−)) heparin rapidly attached to the explant ciliated epithelium in similar numbers. However, only Hep (+) sperm upregulated explant messenger RNA (mRNA) transcription of TLR2, IL8, TGFB1, and PGES, without changes in TNFA and IL-10 expression, while Hep (−) sperm only upregulated PGES. The responses were primarily anti-inflammatory, with a greater response produced by Hep (+) sperm, which also produced a substantial increase in TLR2 protein expression in the epithelium. The addition of TLR1/2 (toll-like receptor 1/2) antagonist to the Hep (+) and (Hep (−)) sperm-explant coincubations reduced sperm attachment to the epithelium and inhibited TLR2 protein expression and some of the Hep (+) sperm-induced mRNA transcription. Our observations suggest that the ampullar epithelium immunologically reacts more strongly to sperm that have undergone heparin stimulation of capacitation. This anti-inflammatory

show abstract

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

Cited by 17 publications

References 29 publications

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

Dynamics of Bovine Sperm Interaction with Epithelium Differ Between Oviductal Isthmus and Ampulla

Toll‐like receptor 2 mediates the immune response of the bovine oviductal ampulla to sperm binding

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