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Home / Equine Research Bank / Animals (Basel) / Effects of In Vitro Interactions of Oviduct Epithelial Cells...
Animals : an open access journal from MDPI2021; 11(1); doi: 10.3390/ani11010074

Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status.

Abstract: Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 × 106/mL compared to lower concentrations (p < 0.05) and when a short centrifugation at 200× g was performed (p < 0.05). Moreover, because sperm suitable for oocyte fertilization is released from oviduct epithelial cells (OECs), in response to the capacitation process, we established an in vitro OEC culture model to select a sperm population with potential fertilizing capacity in this species. We demonstrated E-cadherin and cytokeratin expression in cultures of OECs obtained. When sperm-OEC cocultures were performed, the attached spermatozoa were motile and presented an intact acrosome, suggesting a selection by the oviductal model. When co-cultures were incubated in capacitating conditions a greater number of alive (p < 0.05), capacitated (p < 0.05), with progressive motility (p < 0.05) and with the intact acrosome sperm population was observed (p < 0.05) suggesting that the sperm population released from OECs in vitro presents potential fertilizing capacity. Improvements in handling and selection of cryopreserved sperm would improve efficiencies in ARTs allowing the use of a population of higher-quality sperm.
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Publication Date: 2021-01-03 PubMed ID: 33401609PubMed Central: PMC7823615DOI: 10.3390/ani11010074Google Scholar: Lookup
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Summary

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The study examines how artificial manipulation of frozen-thawed stallion sperm interacts with oviduct epithelial cells in vitro and its effects on their viability, mobility and ability to fertilize an egg (capacitation). The research suggests that improved handling and selection of cryopreserved sperm could enhance assisted reproduction techniques by providing a population of higher-quality, potentially fertilizable sperm.

Study of Stallion Sperm Motility under Different Conditions

  • The researchers first evaluated how the motility of cryopreserved sperm changes under different conditions. These conditions were tested at various sperm concentrations and at varying speeds of centrifugation.
  • The results demonstrated that sperm motility was highest when the sperm were incubated for 30 minutes at a concentration of 30×10 sperm per milliliter.
  • They also found that performing a shorter centrifugation at 200x speed yielded higher motility rates compared to lower speeds/longer times.

Establishing an In Vitro Oviduct Epithelial Cell Culture Model

  • Since sperm which are suitable for fertilizing an oocyte (egg) are released from oviduct epithelial cells (OEC) during the process of capacitation, the researchers established an in vitro culture model of OEC.
  • Through this model, they aimed to select a population of sperm with potential fertilizing capacity specific to this species.
  • The OEC cultures demonstrated E-cadherin and cytokeratin expression, which are markers of healthy epithelial cells.
  • When the sperm were co-cultured with OECs, it was found that the attached sperm were both motile and presented an intact acrosome – the cap-like structure in sperm that houses enzymes needed for fertilization.

Effects on Sperm Capacitation

  • The researchers incubated the above-mentioned co-cultures under conditions favorable for capacitation. They observed a greater number of living, capacitated sperm with progressive motility and an intact acrosome.
  • These results suggested that the sperm released from the OECs in vitro potentially have a higher fertilizing capacity. This implies that the right conditions and methods could help select a higher-quality population of cryopreserved spermatozoa.

Implications for Assisted Reproductive Technologies

  • The study concluded that better management and selection of cryopreserved sperm could increase the efficiency of assisted reproductive technologies.
  • By using sperm populations with a high likelihood of fertilization, these techniques could enable more successful pregnancies, particularly in cases where the use of high-quality sperm is a limiting factor.

Cite This Article

APA
Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. (2021). Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status. Animals (Basel), 11(1). https://doi.org/10.3390/ani11010074

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 11
Issue: 1

Researcher Affiliations

Gimeno, Brenda Florencia
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Bariani, María Victoria
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Laiz-Quiroga, Lucía
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Martínez-León, Eduardo
  • Signaling and Cancer Laboratory, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Inmunología, Genética y Metabolismo, Facultad de Farmacia y Bioquímica, Hospital de Clínicas "José de San Martín", Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires (CABA), CP 1120, Argentina.
Von-Meyeren, Micaela
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Rey, Osvaldo
  • Signaling and Cancer Laboratory, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Inmunología, Genética y Metabolismo, Facultad de Farmacia y Bioquímica, Hospital de Clínicas "José de San Martín", Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires (CABA), CP 1120, Argentina.
Mutto, Adrián Ángel
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.
Osycka-Salut, Claudia Elena
  • Laboratorio de Biotecnologías Reproductivas y Mejoramiento Genético Animal, Instituto de Investigaciones Biotecnológicas, Universidad Nacional de San Martín (UNSAM), Campus Miguelete, Avenida 25 de Mayo y Francia, San Martín, Buenos Aires, CP 1650, Argentina.

Grant Funding

  • PICT 2016-3138 / Fondo para la Investigación Científica y Tecnológica
  • PICT 2015-1548 / Fondo para la Investigación Científica y Tecnológica

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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