Effect of a pre-freezing treatment with cholesterol-loaded cyclodextrins on boar sperm longevity, capacitation dynamics, ability to adhere to porcine oviductal epithelial cells in vitro and DNA fragmentation dynamics

Abstract

The aim of this work was to examine how a pre-freezing treatment with cholesterol-loaded cyclodextrins (CLC) affects boar sperm longevity, capacitation dynamics, ability to bind to a porcine telomerase-immortalised oviductal epithelial cell line (TERT-OPEC) in vitro and DNA integrity dynamics after freeze-thawing. Although the samples treated with CLC exhibited lower sperm quality than the control samples (P<0.05) immediately after thawing, these differences disappeared (P>0.05) after long-term incubation (26h at 37 or 16°C). Additionally, the CLC-treated spermatozoa underwent similar capacitation and DNA fragmentation dynamics as the control spermatozoa (P>0.05). However, CLC-treated spermatozoa were better able to bind to TERT-OPEC in vitro (P<0.0001). In conclusion, the pre-freezing treatment of boar spermatozoa with CLC enhanced the ability of the spermatozoa to bind to TERT-OPEC in vitro, which could have an effect on the establishment of the sperm reservoir in the ampullary--isthmic junction in vivo. Additionally, frozen-thawed spermatozoa can be stored at 16°C for at least 6h without a significant observable decline in sperm quality, which could be beneficial for the transport of thawed diluted doses of spermatozoa from the laboratory to the farm.

Heat shock protein A8 stabilizes the bull sperm plasma membrane during cryopreservation: Effects of breed, protein concentration, and mode of use

“…Subsequently CLCs have been used to improve the freezethawing results in several species, including pig, sheep, stallion, and goat [40][41][42][43][44]. We therefore hypothesized that adding a combination of cholesterol-loaded CLCs and HSPA8 to semen cryopreservation might improve cryopreservation outcomes still further.…”

Recent Advances in Boar Sperm Cryopreservation: State of the Art and Current Perspectives

“…These molecules improve post‐thaw sperm quality and in vitro fertilizing ability without affecting capacitation status and DNA integrity (Tomás et al. , ; Blanch et al. ).…”

“…Another way to improve cryopreservation outcomes is supplementing the media with cholesterol-loaded cyclodextrins (CLC). These molecules improve post-thaw sperm quality and in vitro fertilizing ability without affecting capacitation status and DNA integrity (Tom as et al 2011(Tom as et al , 2013aBlanch et al 2012). Their impact on reproductive performance in vivo is, however, less clear (Tom as et al 2013b).…”

Methods for Improving In Vitro and In Vivo Boar Sperm Fertility

“…Furthermore, although supplementation with cholesterol‐loaded cyclodextrins prior to cryopreservation has been known to improve sperm cryosurvival and acrosome damage (Tomas et al. , ), it may be effective even when spermatozoa are treated with cholesterol‐loaded cyclodextrins in thawing extender (Tomas et al. ).…”

“…Furthermore, although supplementation with cholesterol-loaded cyclodextrins prior to cryopreservation has been known to improve sperm cryosurvival and acrosome damage (Tomas et al 2013, it may be effective even when spermatozoa are treated with cholesterol-loaded cyclodextrins in thawing extender ). The addition of calcium chelators, both EDTA and EGTA, to thawing solution is reported to be a beneficial to improve post-thawed sperm motility and fertility (Okazaki et al 2011).…”