Directional freezing of sperm and associated derived technologies

Arav, Amir; Saragusty, Joseph

doi:10.1016/j.anireprosci.2016.02.007

Cited by 22 publications

(9 citation statements)

References 61 publications

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“…Dried epididymal spermatozoa have never been used in large animal models. Furthermore, in those studies that evaluated DNA fragmentation in lyophilized spermatozoa, only DSBs were evaluated [ 9 , 10 , 22 , 25 , 26 , 27 ].…”

Section: Discussionmentioning

confidence: 99%

DNA fragmentation in epididymal freeze-dried ram spermatozoa impairs embryo development

Palazzese

Gosálvez

Anzalone

et al. 2018

Journal of Reproduction and Development

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Sperm freeze-drying is a revolutionary technique, which has been gaining prominence in recent years. The first related significant result was Wakayama and Yanagimachi’s demonstration in 1998 of the birth of healthy mouse offspring by Intracytoplasmic Sperm Injection (ICSI), using epididymal freeze-dried spermatozoa. Mouse, rat, and hamster models were the first small mammals born from lyophilized epididymal spermatozoa, whereas most other studies in this field used ejaculated spermatozoa. In this work, we applied this technique to ram epididymal spermatozoa, checking the correlation between DNA integrity and embryo development following ICSI. To do this, epididymal sperm from four rams was lyophilized in a trehalose, glucose, KCl, HEPES, and Trolox media. To evaluate DNA damage and fragmentation after rehydration, samples were processed for Sperm Chromatin Dispersion test (SCD), Two-Tailed Comet Assay, and were used for ICSI. Ram #2 had a higher rate of spermatozoa with intact DNA compared with rams #1, #3, and #4 (28% vs. 3.8%, 2.8%, and 5%, respectively) and the lowest rate of Single-Strand Breaks (SSBs) (70% vs. 95.9%, 92.6%, and 93% respectively). Ram #3 had a higher level of Double-Strand Breaks (DSBs) compared to Ram #1 (4.6% vs. 0.33%, respectively). Embryo development to the blastocyst stage following ICSI was only reached from rams whose sperm had higher level of intact DNA – Rams #2 and #4 (6%, 5/147 and 6.3%, 4/64, respectively). Definitively, the impact of sperm DNA damage on embryonic development depends on the balance between sperm DNA fragmentation extent, fragmentation type (SSBs or DSBs), and the oocyte’s repair capacity.

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Section: Discussionmentioning

confidence: 99%

DNA fragmentation in epididymal freeze-dried ram spermatozoa impairs embryo development

Palazzese

Gosálvez

Anzalone

et al. 2018

Journal of Reproduction and Development

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“…However, the formation of blastocysts with normal chromosomes was possible after the transfer of dried nuclei from donor cells into recipient oocytes because of the cytoplast's DNA repair capacity (Iuso et al, 2013; Loi et al, 2008). Overall, DNA and nucleosomal organizations from lyophilized sperm cells contribute to fertilization and embryo development despite overall cellular structural damage and loss of motility (Arav & Saragusty, 2016; Loi et al, 2021). However, there is still a loss of sperm DNA integrity after lyophilization (from 8% to 92% compared to fresh counterparts) in many species studied so far (human [Shahmoradi et al, 2022], cat [Tsujimoto et al, 2020], horse [Restrepo et al, 2019], or sheep [Palazzese et al, 2018]).…”

Section: Damages and Stresses On Biological Structures And Functions ...mentioning

confidence: 99%

Section: Impact On Dna Integrity Epigenetic Factors and Gene Expressionmentioning

confidence: 99%

Damages and stress responses in sperm cells and other germplasms during dehydration and storage at nonfreezing temperatures for fertility preservation

Comizzoli

Amelkina

Lee

2022

Molecular Reproduction Devel

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Long‐term preservation of sperm, oocytes, and gonadal tissues at ambient temperatures has the potential to lower the costs and simplify biobanking in human reproductive medicine, as well as for the management of animal populations. Over the past decades, different dehydration protocols and long‐term storage solutions at nonfreezing temperatures have been explored, mainly for mammalian sperm cells. Oocytes and gonadal tissues are more challenging to dehydrate so little to no progress have been made. Currently, the detrimental effects of the drying process itself are better characterized than the impact of long‐term storage at nonfreezing temperatures. While structural and functional properties of germ cells can be preserved after dehydration, a long list of damages and stresses in nuclei, organelles, and cytoplasmic membranes have been reported and sometimes mitigated. Characterizing those damages and better understanding the response of germ cells and tissues to the stress of dehydration is fundamental. It will contribute to the development of optimal protocols while proving the safety of alternative storage options for fertility preservation. The objective of this review is to (1) document the types of damages and stress responses, as well as their mitigation in cells dried with different techniques, and (2) propose new research directions.

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“…The propagation of the samples on the cold blocks and the gap will be what defines the cooling rate. This process of directional freezing generates the continuous “seeding” previously mentioned , producing a controlled nucleation of ice throughout the process, which is extremely beneficial since it avoids supercooling, the abrupt release of post‐freezing latent heat and the formation of ice crystals (Arav & Saragusty, ; Arav et al, ) .…”

Section: Basic Principles Of Cryopreservationmentioning

confidence: 99%

Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post‐thaw quality optimization

Grötter

Cattáneo

Marini

et al. 2019

Reprod Domestic Animals

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Contents Sperm cryopreservation facilitates the storage and transport of germplasm for its use in artificial insemination (AI) and other advanced reproductive technologies. The cryopreservation process can damage sperm and compromise functionality. Several cryobiological studies have found that the physical and biological factors that affect sperm survival at low temperatures during the cryopreservation process often involve the integrity of sperm membrane. In this review, the behaviour of the sperm membrane against cooling, cold shock, ice crystal formation, oxidative stress, osmotic changes, reorganization of the lipid bilayer and addition of cryoprotective agents (CPA) is discussed. In addition, the phenomenon of reactive oxygen species (ROS) and its relationship with the cryopreservation process is also described. Semen cryopreservation techniques have progressed slowly in past years, and the current performance, measured as post‐thawed survival, is not very different compared to past decades. Recent advances in understanding the structure of the cell membrane, its function and metabolism have driven to new conservation systems, including lyophilization and vitrification. However, none of these technologies is commercially available, although its future appears very promising.

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Directional freezing of sperm and associated derived technologies

Cited by 22 publications

References 61 publications

DNA fragmentation in epididymal freeze-dried ram spermatozoa impairs embryo development

DNA fragmentation in epididymal freeze-dried ram spermatozoa impairs embryo development

Damages and stress responses in sperm cells and other germplasms during dehydration and storage at nonfreezing temperatures for fertility preservation

Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post‐thaw quality optimization

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