Characteristics and fertility of domestic cat epididymal spermatozoa cryopreserved with two different freezing media

Brusentsev, E. Yu.; Kizilova, Elena A.; Mokrousova, Valentina; Kozhevnikova, V. V.; Рожкова, И. Н.; Amstislavsky, S. Ya.

doi:10.1016/j.theriogenology.2017.12.038

Cited by 14 publications

(6 citation statements)

References 23 publications

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“…The length of storage time did not significantly influence oocyte activation, pronuclear formation, or embryo development. This is also similar to the outcome of cryopreservation for feline spermatozoa [43,44]. Post-ICSI oocyte activation by ethanol was required to overcome fertilization failure in the current system; however, alternative activation methods should be explored in the future to lower the rate of parthenogenesis [45,46].…”

Section: Discussionmentioning

confidence: 52%

Desiccated cat spermatozoa retain DNA integrity and developmental potential after prolonged storage and shipping at non-cryogenic temperatures

Lee

Zahmel

Jewgenow

et al. 2021

J Assist Reprod Genet

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Purpose To evaluate the DNA integrity and developmental potential of microwave-dehydrated cat spermatozoa after storage at − 20 °C for different time periods and/or overnight shipping on dry ice. Methods Epididymal spermatozoa from domestic cats were microwave-dehydrated on coverslips after trehalose exposure. Dried samples were either assessed immediately, stored for various duration at − 20 °C, or shipped internationally on dry ice before continued storage. Dry-stored spermatozoa were rehydrated before assessing DNA integrity (TUNEL assays) or developmental potential (injection into in vitro matured oocytes followed by in vitro embryo culture for up to 7 days). Results Percentages of dried-rehydrated spermatozoa with intact DNA was not significantly affected (P > 0.05) by desiccation and short-term storage (range, 78.9 to 80.0%) but decreased (P < 0.05) with storage over 5 months (range, 71.0 to 75.2%) compared to fresh controls (92.6 ± 2.2%). After oocyte injection with fresh or dried-rehydrated spermatozoa (regardless of storage time), percentages of activation, pronuclear formation, and embryo development were similar (P > 0.05). Importantly, spermatozoa shipped internationally also retained the ability to support embryo development up to the morula stage. Conclusion Results demonstrated the possibility to sustain DNA integrity and developmental potential of spermatozoa by dry-preservation, even after long-term storage and long-distance shipment at non-cryogenic temperatures. While further studies are warranted, present results demonstrate that dry preservation can be a reliable approach for simple and cost-effective sperm biobanking or shipment.

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Section: Discussionmentioning

confidence: 52%

Desiccated cat spermatozoa retain DNA integrity and developmental potential after prolonged storage and shipping at non-cryogenic temperatures

Lee

Zahmel

Jewgenow

et al. 2021

J Assist Reprod Genet

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“…In addition, greater sensitivity of feline epididymal sperm to cryoinjuries was inferred from this result (Hermansson & Axnér, 2007). The literature shows sperm motility values after thawing from 15.5% to 40% for cat epididymal sperm (Macente et al, 2012;Buranaamnuay, 2015;Kunkitti et al, 2016;Prochowska et al, 2016;Brusentsev et al, 2018); and 52.8% -70.6% for semen from the same species (Platz et al, 1978;Villaverde, 2007;Chatdarong et al, 2010).…”

Section: Discussionmentioning

confidence: 76%

Brazilian green propolis doesn't have a beneficial effect on the cryopreservation of domestic cat epididymal spermatozoa

Gomes

Fernandes

Izzo

et al. 2021

RSD

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This study evaluated the effect of different concentrations of Brazilian green propolis on the cryopreservation of epididymal sperm from domestic cats. Spermatozoa were collected from cauda epididymis by slicing technique in a TRIS extender, preheated to 37ºC. Then, the sperm were evaluated and divided randomly between the control group (without adding propolis in the extender) and the treatment groups (adding different concentrations of propolis extract in the extender): P1 (0.1 mg/mL), P2 (0.3 mg/mL) e P3 (0.6 mg/mL). Subsequently, sperm were cryopreserved. The evaluated parameters were sperm kinetics by the Computer Assisted Semen Analysis, vigor, viability, membrane functionality, chromatin condensation and morphology. The parameters were measured before and after cryopreservation. Propolis didn’t show toxicity to the sperm in any concentrations, with no changes observed in the motility pattern. There was no significant influence of propolis on motility, vigor, percentage of viable spermatozoa, chromatin quality and other kinetic parameters in the cryopreserved samples. In sperm morphology, it was demonstrated a reduction in the percentage of normal cells in P1 and P2 groups in relation to the fresh sample, control and P3 group. In addition, an interesting effect from the propolis groups was observed on possible growth inhibition of microorganisms in the contaminated samples. Propolis didn’t provide superiority to the sperm parameters evaluated after thawing, except for the values of plasma membrane functionality, which were better. The propolis is an interesting component to be incorporated in the cryoprotectant medium due to its non-toxicity and the potential inhibition of microbial growth.

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“…Furthermore, the process of cryopreservation uses molecules such as cryoprotectants that can be toxic to spermatozoa, which makes colling and freezing semen more challenging (Cheuquemán et al., 2018; Pukazhenthi et al., 1999; Souza et al., 2018). Therefore, different extenders are being tested to improve the cryopreservation results for these species (Brusentsev et al., 2018).…”

Section: Introductionmentioning

confidence: 99%

Insights into alternative cryoprotectants to freeze sperm of domestic cats

Martins,

Silva,

Hidalgo

et al. 2024

Reprod Domestic Animals

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Globalization and habitat destruction pose a significant threat to wildlife felids. Even though conservation banks for genetic materials have been created, the sperm cryopreservation with minimal cell damage is still a great challenge. Thus, this study aimed to compare the effects of two commercial extenders with different concentrations of alternative cryoprotectants on thawed sperm quality of domestic cats. Five adult cats were anaesthetized (using a combination of 40 μg/kg medetomidine associated to 5 mg/kg ketamine), and the semen was collected by electroejaculation (electrical stimulation of 2–3 V). Semen samples were evaluated for sperm characteristics (kinetics, morphology, membrane integrity and morphometry). Subsequently, they were sorted into two aliquots and centrifuged. The aliquots were added to a commercial extender containing 3% glycerol and 2% methylformamide (extender I) or 2% glycerol and 3% methylformamide (extender II), frozen, thawed (37°C/30 s) and reevaluated. Comparatively, the sperm kinetics and membrane integrity of fresh semen were higher (p < .002) than frozen samples in extender I and II. Total and progressive motility were lowest in the thawed samples. However, the subjective analysis indicated high sperm motility, since the kinetics evaluation was impaired by the low cell number in the thawed samples. There were no differences in sperm morphology between the groups. In the sperm morphometric analysis, a significant difference (p = .04) was identified in the length of the intermediate piece in extender II samples compared with fresh and extender I. Thus, it can be concluded that although the concentrations tested did not maintain the kinetic parameters and membrane integrity of spermatozoa after thawing, the extender with a lower concentration of glycerol was less toxic for maintaining the midpiece length.

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Characteristics and fertility of domestic cat epididymal spermatozoa cryopreserved with two different freezing media

Cited by 14 publications

References 23 publications

Desiccated cat spermatozoa retain DNA integrity and developmental potential after prolonged storage and shipping at non-cryogenic temperatures

Desiccated cat spermatozoa retain DNA integrity and developmental potential after prolonged storage and shipping at non-cryogenic temperatures

Brazilian green propolis doesn't have a beneficial effect on the cryopreservation of domestic cat epididymal spermatozoa

Insights into alternative cryoprotectants to freeze sperm of domestic cats

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