Minimal volume vitrification of epididymal spermatozoa results in successful in vitro fertilization and embryo development in mice

Abstract

This study compared three cryopreservation protocols on sperm functions, IVF outcomes, and embryo development. Epididymal spermatozoa cryopreserved using slow-cooling (18% w/v raffinose, RS-C) were compared with spermatozoa vitrified using 0.25 M sucrose (SV) or 18% w/v raffinose (RV). The motility, vitality, and DNA damage (TUNEL assay) of fresh control (FC) spermatozoa were compared with post-thawed or warmed RS-C, RV, and SV samples. Mouse oocytes (n = 267) were randomly assigned into three groups for insemination: RV (n = 102), RS-C (n = 86), and FC (n = 79). The number and the proportion of two-cell embryos and blastocysts from each treatment were assessed. Sperm motility (P < 0.01) and vitality (P < 0.05) were significantly reduced after vitrification compared with slow-cooled spermatozoa. However, DNA fragmentation was significantly reduced in spermatozoa vitrified using sucrose (15 ± 1.8% [SV] vs 26 ± 2.8% [RV] and 27 ± 1.2% [RS-C]; P < 0.01). Although the number of two-cell embryos produced by RS-C, RV, and FC spermatozoa was not significantly different, the number of blastocysts produced from two-cell embryos using RV spermatozoa was significantly higher than FC spermatozoa (P = 0.0053). This simple, small volume vitrification protocol and standard insemination method allows successful embryo production from small numbers of epididymal spermatozoa and may be applied clinically to circumvent the need for ICSI, which has the disadvantage of bypassing sperm selection.

Human sperm parameter improvement associated with Ceratonia siliqua extract as a cryopreservation supplement after vitrification

“…One method to prevent the formation of ice crystals and the associated damage is cryopreservation by vitrification [ 30 ]. In this process, sperm cryopreservation is performed more quickly and with greater safety than in other methods [ 31 , 32 ]. The cryopreservation process plays a role in the production of ROS by accelerating the conversion of anion superoxide to hydrogen peroxide [ 33 ].…”

Effect of aqueous Nigella sativa extract on the functional parameters of post-thaw human spermatozoa during vitrification

“…Vitrification is a relatively new method for sperm cryopreservation that has advantages over conventional methods, such as reducing intracellular ice crystals. Moreover, compared to slow freezing, vitrification is cheaper and freezes sperm in less time [ 5 ]. Sperm naturally contains many antioxidant systems capable of reducing intracellular damage [ 6 ].…”

The protective effect of zinc oxide and selenium oxide nanoparticles on the functional parameters of rat sperm during vitrification

“…The vitrification method is an effective technique for sperm freezing, as it prevents the formation of ice crystals [ 3 ]. This method is safer, less expensive, and quicker than other sperm freezing methods [ 4 , 5 ]. Regardless of the method used, the sperm freezing process can negatively impact sperm quality due to the excessive production of free oxygen radicals and subsequent oxidative stress.…”

“…Freezing is a technique used to preserve fertility in young men with cancer who are candidates for chemotherapy and radiation er, less expensive, and quicker than other sperm freezing methods [4,5]. Regardless of the method used, the sperm freezing process can negatively impact sperm quality due to the excessive production of free oxygen radicals and subsequent oxidative stress.…”