Cryopreservation of horse semen with a liposome and trehalose added extender

Abstract

The aim of the study was to evaluate the effect of cryopreserved equine semen in the presence of trehalose-loaded liposomes on the integrity and function of sperm cells. Six ejaculations of five stallions collected with an artificial vagina were used. The ejaculates were diluted with INRA 96 ® 2:1 v/v and transported at 22 °C to the laboratory. Before cryopreservation, the semen was diluted with INRA Freeze ® to obtain the following treatments: T1) INRA Freeze ® (control), T2) INRA Freeze ® + liposomes, T3) INRA Freeze ® + liposomes+trehalose. Data were analysed using the Kruskal Wallis test. The percentages of sperm with intact DNA were 54.5, 57.9, and 64.8% for T1, T2 and T3, respectively (P>0.05). When evaluating the acrosomal and capacitation state after filtering with Percoll ® , the percentages of spermatozoa without acrosome reaction and without capacitation were 67.8, 79.2 and 68.1% in T1, T2, and T3, respectively (P>0.05), while the capacitated sperm without acrosome reaction and without capacitation was similar in T1 (47%) and T3 (32%) (P>0.05), and lower in T2 (16%) before filtering with Percoll ®. The use of liposomes and liposome-trehalose did not affect on the functional status and nuclear chromatin of the equine sperm after freezing, but it did affect the percentage of capacitated sperm without acrosome reaction after selecting the thawed semen using the Percoll ® gradient.

Sperm Cryopreservation in Farm Animals Using Nanotechnology

“…The primary aim for sublime sperm functionality is to provide optimum pH, temperature, and osmolarity. In humans, cryoprotective media reduce ice crystal formation and cold shock, resulting in sperm functionality [80]. Permeable and non-permeable cryoprotectants have 4-5% glycerol and 20% egg yolk that guard against cryodamage.…”

Effect of Sperm Cryopreservation in Farm Animals Using Nanotechnology

“…Although some literature has proven the positive effects of soy lecithin (vegetable origin), egg yolk (animal origin) is still used as cryopreservation outcomes vary significantly. Liposomes are free from contamination and are used as an alternative to freezing media in sperm cryopreservation of equine, swine and bovine families [ 79 , 80 ]. The plasma membrane of liposomes controls the permeability of water and cryoprotectants [ 81 ].…”

The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation

“…Moreover, liposomes can be loaded with lipid-related content (such as lecithin [110]) to improve the plasma membrane regeneration efficacy during the freeze-thaw process of ram spermatozoa. Liposomes were used as a cryoprotectant additives in several animal species including equine [111,112], buffalo [113], ovine [107,114,115], porcine [116], and bovine [117] with reported improvement in fertility after AI [118]. It has been proposed that liposomes with their contents of phospholipids (phosphatidylserine, dioleoylphosphatidylcholine, phosphatidylcholine, dipalmitoylphosphatidylcholine, and dimyristoylphosphocholine) and saturated and unsaturated fatty acids can fuse with the sperm plasma membrane and abate the damage to spermatozoa caused by the freeze-thaw process [119,120] (Figure 1).…”