Effect of different carboxylated poly l-lysine and dimethyl sulfoxide combinations on post thaw rabbit sperm functionality and fertility

Küçük, Niyazi; Raza, Sanan; Matsumura, Kazuaki; Uçan, Uğur; Serin, İlker; Ceylan, Ahmet; Aksoy, Melih

doi:10.1016/j.cryobiol.2021.07.001

Cited by 9 publications

(13 citation statements)

References 38 publications

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“…The most suitable concentration of CPLL in the presence of 3% glycerol for pig spermatozoa was 0.25%, whereas the most suitable concentration of CPLL in the presence of 3.25% glycerol for bovine spermatozoa was 0.5% [ 11 ]. Moreover, the most suitable concentrations of CPLL have been reported to be 0.75% in the presence of 5% glycerol for buffalo spermatozoa [ 12 ], 0.3% in the presence of 7% glycerol for human spermatozoa [ 14 ] and 1% in the presence of 8% DMSO for rabbit spermatozoa [ 13 ]. These results suggest that the optimal concentration of CPLL for cryopreservation of spermatozoa varies with the species.…”

Section: Discussionmentioning

confidence: 99%

“…The reduction in the cellular stress may be responsible for cryoprotection [ 9 ]. Recent studies have shown that CPLL is also effective for cryopreservation of germ cells such as pig embryos at the pronuclear stage [ 10 ] and bovine, buffalo, rabbit, and human spermatozoa [ 11 , 12 , 13 , 14 ]. In the present study, we investigated the efficacy of CPLL as a cryoprotectant for pig spermatozoa.…”

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confidence: 99%

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Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant

JIN

Choi

Matsumura

et al. 2022

Journal of Reproduction and Development

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In this study, we cryopreserved pig spermatozoa using carboxylated poly-L-lysine (CPLL) as the cryoprotectant to determine its efficacy. Pig spermatozoa were placed in a freezing extender containing 3% (v/v) glycerol and different CPLL concentrations. The motility indices of the spermatozoa cryopreserved with 0.25% (v/v) CPLL at 6 (59.3), 9 (53.7), and 12 (26.2) h after thawing were significantly higher (P < 0.01 or P < 0.05) than those of the spermatozoa cryopreserved without CPLL (53.7, 40.1, and 17.5 at 6, 9, and 12 h after thawing, respectively). The concentration of CPLL in the freezing extender did not affect the ability of frozen-thawed spermatozoa to fertilize oocytes in vitro . However, the blastocyst formation rate of embryos derived from spermatozoa cryopreserved with 0.25% CPLL (24.6%) was significantly higher (P < 0.01) than that of embryos derived from spermatozoa cryopreserved without CPLL (11.2%). The conception rate of the sows inseminated with spermatozoa cryopreserved with 0.25% CPLL (72.2%) was not significantly different from that of the sows inseminated with spermatozoa stored at 17°C (81.3%). However, the mean number of total piglets born to the former (10.0) was significantly lower (P < 0.05) than that of total piglets born to the latter (13.4). The results showed that CPLL in the freezing extender maintained the motility of frozen-thawed pig spermatozoa and improved the in vitro development of embryos produced by in vitro fertilization. In addition, we have demonstrated that piglets could be obtained with artificial insemination using spermatozoa cryopreserved with CPLL.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant

JIN

Choi

Matsumura

et al. 2022

Journal of Reproduction and Development

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“…Frozen semen was stored in a liquid nitrogen tank for 2 months (Salisbury et al 1978). Then, frozen semen samples were thawed at 37°C for 30 s (Küçük et al 2021). Sperm analyzes were performed with CASA in thawed semen samples (Castellini et al 2011).…”

Section: Animals Semen Collection Cryopreservation Process and Sperm ...mentioning

confidence: 99%

The effect of luteolin on spermatological parameters, apoptosis, oxidative stress rate in freezing rabbit semen

Akarsu

Acısu

Güngör

et al. 2023

Polish Journal of Veterinary Sciences

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The aim of the present study was to determine the effects of Luteolin (LUT) on semen quality, oxidative stress, apoptosis, acrosomal integrity, mitochondrial membrane potential and dead sperm ratio in rabbits. Ejaculates from six New Zealand rabbits were collected, evaluated and pooled. The pooling was divided into five groups as control (no additive) LUT 25 µM, LUT 50 µM, LUT 100 µM and LUT 200 µM and LUT added. It was then filled into a falcon tube with Tris-based extender at a final concentration of approximately 35 x 10 6 spermatozoa. Diluated rabbit semen samples were drawn into frozen and thawed. Frozen semen straws were thawed at 37°C in 30 seconds. According to our findings, no statistical difference was found between all doses of luteolin and the control group in the CASA (computer assisted sperm analysis) analysis performed at 4°C. However, total motility, progressive motility and rapid sperm percentage were found to be higher in the frozen and thawed rabbit semen at a dose of LUT 50 µM compared to the other groups (p<0.05). While amplitude of lateral head displacement (ALH) and beat cross-frequency (BCF) values were found at the lowest dose of LUT 200 µM, a statistically significant difference was observed between the other groups. When the flow cytometry results were examined, no statistical difference was found between the rate of dead sperm, acrosomal integrity, mitochondrial membrane potential and apoptosis rate. Morever, the H 2 O 2 percentage was found to be lower in all experimental groups compared to the control group (p<0.001). In conclusion, the addition of LUT in long-term storage of rabbit semen provided a protective effect for spermatozoa with its antioxidative properties against damage caused by cryopreservation.

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“…Eventually, the cooled straws were frozen with liquid nitrogen vapor 6-7 cm above the surface for 10 min, just before directly plunging into the liquid nitrogen. 11 The frozen sperm samples were stored in liquid nitrogen until the start of sperm analyses.…”

Section: Animals Sperm Collection and Cryopreservationmentioning

confidence: 99%

“…The sperm viability and acrosome integrity were assessed with propidium iodide (PI) and Lectin from Arachis hypogaea (peanut) fluorescein isothiocyanate conjugate (FITC-PNA) staining procedure. 11 Briefly, 50 μL sperm sample was incubated and stained with 2.5 μL PI (500 μg/ mL) and 5 μL FITC-PNA (200 μg/mL) for 10 min at room temperature. The stained 3 μL sperm sample was placed on slide and covered with cover slip.…”

Section: Evaluation Of Sperm Viability and Acrosome Integritymentioning

confidence: 99%

Effect of Extenders Including High Concentrations Dimethyl Sulfoxide (DMSO) on Post-Thaw Rabbit Sperm Parameters

Küçük

2022

Journal of Research in Veterinary Medicine

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Cryoprotectants have critical roles to prevent cell damages during cryopreservation. However, the adjustment of cryoprotectant concentration is also very crucial to protect cells from cryoprotectant toxicity. The present study was designed to investigate the effect of extenders including high concentration dimethyl sulfoxide (DMSO) on post-thaw rabbit sperm quality. Pooled rabbit sperm samples (n=7) were diluted and cryopreserved in extenders including 250 mmol/L Tris, 88 mmol/L citric acid, 47 mmol/L glucose, 1% sucrose and different concentrations of DMSO. The presence of high concentration DMSO (12 and 14%) in extender decreased sperm total and progressive motility (P < 0.01). The 8 and 10 percent DMSO supplementations in extender increased live sperm rates (P < 0.01). Live and intact acrosome or intact membrane sperm rates were detected higher in 8 and 10 DMSO groups (P < 0.05). When total intact membrane sperm rates were similar in all groups, total intact acrosome sperm rate was higher in 8 DMSO group compared to 12 DMSO group (P < 0.05). In conclusion, when 12 and 14 percent DMSO additions in extender adversely affected post-thaw sperm parameters, the presence of 8 percent DMSO in extender provided the highest post-thaw sperm quality.

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Effect of different carboxylated poly l-lysine and dimethyl sulfoxide combinations on post thaw rabbit sperm functionality and fertility

Cited by 9 publications

References 38 publications

Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant

Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant

The effect of luteolin on spermatological parameters, apoptosis, oxidative stress rate in freezing rabbit semen

Effect of Extenders Including High Concentrations Dimethyl Sulfoxide (DMSO) on Post-Thaw Rabbit Sperm Parameters

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