97 Improved Cryopreservation of Domestic Cat Spermatozoa in a Soy Lecithin-Based Extender

Abstract

Development of a chemically defined, plant-based cryopreservation media would reduce extender variability and the potential for transmission of zoonotic pathogens compared with traditional egg-yolk-based extenders. The objective of this study was to compare effects of egg yolk- and soy lecithin-based cryopreservation media and the temperature of glycerol addition on sperm parameters following freezing and thawing of domestic cat spermatozoa. Fresh semen was collected by manual stimulation on 3 separate occasions from 4 adult male cats. Each ejaculate was washed to remove seminal plasma, divided into 4 equal aliquots, and extended at room temperature in one of the following treatments: 1) TEST-egg yolk (Irvine Scientific Inc., Santa Ana, CA, USA) medium with 4% glycerol (EYG); 2) TEST-egg yolk, with 4% glycerol added after cooling to 5°C (EY); 3) TES-Tris buffer with soy-lecithin (1%) and 4% glycerol (SLG); and 4) TES-Tris buffer with 1% soy-lecithin, and 4% glycerol added after cooling to 5°C (SL). Sperm progressive motility (%) and rate of progressive movement (scale of 0–5) were evaluated at 0, 1, 3, 6, and 24 h post-thaw. Sperm capacitation (chlortetracycline staining), acrosome integrity (FITC-PNA staining), and DNA integrity (acridine orange staining) were assessed at 15 min post-thaw. Data were exponentially transformed to achieve normal distribution and then subjected to GLM analysis to determine effects of media and temperature of glycerol addition on sperm traits. At 0 and 1 h post-thaw, acrosome integrity, DNA integrity and % sperm motility did not differ (P > 0.05) among treatments. However, % sperm motility was greater in the soy-based media compared to egg yolk-based media at 3, 6, and 24 h post-thaw (Table 1; P < 0.05). A higher percentage of uncapacitated spermatozoa were present in soy-based compared to egg-yolk based cryopreservation media (63.9 ± 9.3 v. 51.2 ± 11.5, respectively; P < 0.05), regardless of temperature of glycerol addition. Finally, addition of glycerol at 5°C resulted in higher % sperm motility compared to room temperature at 6 and 24 h post-thaw in both medium types (Table 1; P < 0.05). Our results suggest that use of a chemically defined, soy-based medium improves long-term motility and capacitation status of frozen–thawed domestic cat spermatozoa compared with cryopreservation in a traditional egg yolk-based extender. Table 1.Motile spermatazoa and motility score at 3, 6, and 24 h

Ram semen preserved at 0°C with soybean lecithin Tris-based extender substituted for egg yolk

“…The current study revealed that 0.5% of SL can replace the egg yolk in the Tris dilution, so that the sheep sperm can be stored at 0°C for a longer time. It can be concluded from the literature that the optimal concentration range of SL for cryopreservation is 0.8% in dogs [ 20 ] to 1% in rams [ 14 ], humans [ 21 ] and cats [ 22 ] and goats [ 23 ]. The reason for this difference in addition of SL is that the differences in species are related to the differences of sperm plasma membrane composition between species [ 24 ].…”

Lecithin nanoparticles enhance the cryosurvival of caprine sperm

“…Soy lecithin, mainly composed of phosphatidylcholine [6], emerged as an alternative to components of animal origin. Many studies have confirmed the suitability of soy lecithin on sperm, including goat [7][8][9][10], ram [11][12][13], human [14] and cat [15], and several commercial formulations are currently available (e.g., Andromed, BIOXCell).…”

Effect of Propolis Ethanolic Extract Supplementation to Ram Semen Extenders on Sperm Characteristics, Lipid Peroxidation and some Enzymatic Activities in Seminal Plasma in Chilled Semen

“…Lecithin is a valuable plant origin to prevent sperm cold shock, because it consists of glycerol, two fatty acids, phosphate group and choline which play an important role in protecting sperm cell membrane. The previous facts has been developed and utilized to preserve spermatozoa of human (Jeyendran et al, 2008), domestic cat (Vick et al, 2010), stallion (Papa et al, 2011), ram (Khalifa and Abdel-Hafez, 2014), buffalo (Akhter et al, 2012), goat (Chelucci et al, 2015) and bovine bull (El-Sisy et al, 2016).…”