Prolonged incubation of frozen–thawed equine spermatozoa for in vitro fertilization: A preliminary study using low temperature and INRA96 medium

• Published in: Reproduction in domestic animals Vol. 59; no. S3; pp. e14593 - n/a
• Main Authors: Luis‐Calero, Marcos, Muñoz‐García, Carmen C., Fernández‐Hernández, Pablo, Macías‐García, Beatriz, González‐Fernández, Lauro
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.10.2024
• Subjects: Acrosome; Animals; Cell Survival; Cold Temperature; Cryopreservation - veterinary; Culture Media; Epinephrine - pharmacology; equine; Fertilization in Vitro - veterinary; frozen–thawed spermatozoa; Horses; Horses - physiology; In vitro fertilization; Incubation; Integrity; Low temperature; Male; Motility; Semen; Semen Preservation - methods; Semen Preservation - veterinary; Sperm; Sperm Motility; Spermatozoa; Spermatozoa - physiology; Viability; motility; frozen–thawed spermatozoa; equine; viability
• ISSN: 0936-6768; 1439-0531; 1439-0531
• DOI: 10.1111/rda.14593

A protocol for conventional in vitro fertilization (IVF) in horses using fresh semen has been described, using a prolonged incubation in FERT‐TALP medium (22 h) at 38.2°C in the presence of penicillamine, hypotaurine and epinephrine (PHE). Our work aimed to develop a protocol that maintains quality parameters in frozen–thawed equine spermatozoa incubated for 22 h in the presence of PHE using different media (FERT‐TALP and INRA96) and incubation temperatures (30 and 38.2°C). Twelve frozen ejaculates from four stallions were thawed and then incubated in either FERT‐TALP or INRA96 with PHE at 30 or 38.2°C for 22 h. Following incubation, total motility (TM), progressive motility (PM), viability and acrosome integrity were evaluated. The results showed that TM was significantly higher (p < .001) at 30°C in both media, while PM was higher for INRA96 at 30°C compared to 38°C (p < .05). Moreover, INRA96 at 30°C exhibited higher sperm viability and acrosome integrity (p < .001) compared to the other experimental groups. These preliminary results suggest that incubating thawed equine spermatozoa at 30°C with PHE in INRA96 successfully maintains motility, viability and acrosome integrity in equine spermatozoa, indicating its potential use for conventional equine IVF.