Chromosomal integrity and DNA damage in freeze‐dried spermatozoa

Kusakabe, Hirokazu

doi:10.1007/s12522-011-0092-7

Cited by 7 publications

(6 citation statements)

References 122 publications

(153 reference statements)

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“…3c ). Results of this study and previous reports 17 , 18 , 20 – 24 suggest that cryoprotectant protect the cell organelle and not the DNA from mechanical stress. Therefore, if our method (maintaining vacuum in ampoules) is combined with using cryoprotectant, morphological integrity as well as DNA integrity of FD spermatozoa will be protected, thereby increasing the success rate of the offspring from those spermatozoa.…”

Section: Discussionsupporting

confidence: 77%

“…For example, Tardigrades can survive under extreme dehydration conditions because of the accumulation of large amounts of trehalose in their bodies which maintains the integrity of cell membranes or organelles 19 . However, it has been reported that cryoprotectants increase the birth rate from dried spermatozoa as well as the success rate of somatic cell cloning 17 , 18 , 20 – 24 , spermatozoa and cells used in these studies were no longer viable following the drying treatment even in the presence of trehalose, and the cell membranes were largely damaged 3 . However, it may be argued that preserving the integrity of the DNA of FD spermatozoa is more important than maintaining the viability of sperm.…”

Section: Discussionmentioning

confidence: 90%

“…Spermatozoa preserved by evaporative drying in a LiCl sorption jar in the presence of trehalose, as a cryoprotectant, have been used to obtain healthy offspring after storage at ambient temperatures for two years 17 . Although FD spermatozoa can be preserved in easy to handle glass ampoules, the freeze-drying machine is expensive, therefore it was unclear which of the two methods, evaporative drying or freeze-drying, was superior for preserving mammalian spermatozoa at RT 18 .…”

Section: Discussionmentioning

confidence: 99%

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Assessing the tolerance to room temperature and viability of freeze-dried mice spermatozoa over long-term storage at room temperature under vacuum

Kamada

Wakayama

Shibasaki

et al. 2018

Sci Rep

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Freeze-drying has been frequently used to preserve food and microorganisms at room temperature (RT) for extended periods of time; however, its application to mammalian species is difficult. Here, we developed a method to prolong the stability of freeze-dried (FD) mice spermatozoa at RT for more than one year without using any cryoprotectant agents. Our data showed that maintaining a vacuum in ampoules is critical to ensuring the viability of FD spermatozoa, as the stability of spermatozoa DNA increased when imperfectly vacuumed ampoules were detected using a non-destructive test and eliminated. Finally a large number of healthy offspring were obtained from mice oocytes fertilized with FD spermatozoa stored at RT for more than one year. Although the birth rate from three-month stored spermatozoa was lower than that from one-day stored spermatozoa, no further reduction was observed even in one-year stored spermatozoa. Therefore, FD spermatozoa preserved in this study were highly tolerant to warm temperatures. This method of storage shows a great potential for the preservation of genetic resources of mammalian species, such as genetically-modified mouse strains, without the use of electric power.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

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Assessing the tolerance to room temperature and viability of freeze-dried mice spermatozoa over long-term storage at room temperature under vacuum

Kamada

Wakayama

Shibasaki

et al. 2018

Sci Rep

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“…However, the birth rate of embryos fertilized with FD sperm was largely decreased compared with that of embryos fertilized with fresh spermatozoa due to DNA damage [29]. Kusakabe suggested that DNA damage was induced by mechanical or oxidative stress not only during freeze-drying but also during long-term preservation [30]. Kaneko et al .…”

Section: Discussionmentioning

confidence: 99%

Effect of trehalose on the preservation of freeze-dried mice spermatozoa at room temperature

Ito

Wakayama

Kamada

et al. 2019

Journal of Reproduction and Development

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Freeze-drying of spermatozoa is a convenient and safe method to preserve mammalian genetic material without the use of liquid nitrogen or a deep freezer. However, freeze-dried spermatozoa (FD sperm) are not frequently used because of the low success rate of offspring after intracytoplasmic spermatozoa injection (ICSI). In this study, we determined the optimal concentration and a point of action of trehalose as a protectant for the preservation of FD sperm from different mouse strains at room temperature (RT). Although trehalose demonstrated no potential to protect the FD sperm of ICR mice against the freeze-drying procedure itself, the blastocyst rate was significantly improved when FD sperm was preserved for more than 1 month at RT (56–63% vs. 29% without trehalose). The optimal concentration of trehalose was 0.5 M. Importantly, remarkable results were obtained when spermatozoa of inbred mouse strains (C57BL/6N, C3H/He, and 129/Sv) were used, and many offspring were obtained when FD sperm that was preserved for 3 months at RT (26–28% vs. 6–11% of without trehalose) was used. However, when DNA damage in FD sperm was examined by gamma-H2Ax assays, it was found that trehalose failed to protect the FD sperm from DNA damage. These results suggest that trehalose has the potential to protect other sperm factors rather than sperm DNA during preservation at RT for longer periods and trehalose is more effective for inbred mouse strains.

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“…) [ 83 , 84 , 85 ]. Furthermore, short-term storage [ 86 ] and worldwide transportation at ambient temperatures [ 87 ] are also possible. A similar simple sperm preservation method using evaporation has been investigated in the mouse [ 88 ].…”

Section: Simple Gamete Preservation By Freeze-drying Spermmentioning

confidence: 99%

Reproductive technologies for the generation and maintenance of valuable animal strains

Kaneko

2018

Journal of Reproduction and Development

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Many types of mutant and genetically engineered strains have been produced in various animal species. Their numbers have dramatically increased in recent years, with new strains being rapidly produced using genome editing techniques. In the rat, it has been difficult to produce knockout and knock-in strains because the establishment of stem cells has been insufficient. However, a large number of knockout and knock-in strains can currently be produced using genome editing techniques, including zinc-finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) system. Microinjection technique has also contributed widely to the production of various kinds of genome edited animal strains. A novel electroporation method, the “Technique for Animal Knockout system by Electroporation (TAKE)” method, is a simple and highly efficient tool that has accelerated the production of new strains. Gamete preservation is extremely useful for maintaining large numbers of these valuable strains as genetic resources in the long term. These reproductive technologies, including microinjection, TAKE method, and gamete preservation, strongly support biomedical research and the bio-resource banking of animal models. In this review, we introduce the latest reproductive technologies used for the production of genetically engineered animals, especially rats, using genome editing techniques and the efficient maintenance of valuable strains as genetic resources. These technologies can also be applied to other laboratory animals, including mice, and domestic and wild animal species.

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Chromosomal integrity and DNA damage in freeze‐dried spermatozoa

Cited by 7 publications

References 122 publications

Assessing the tolerance to room temperature and viability of freeze-dried mice spermatozoa over long-term storage at room temperature under vacuum

Assessing the tolerance to room temperature and viability of freeze-dried mice spermatozoa over long-term storage at room temperature under vacuum

Effect of trehalose on the preservation of freeze-dried mice spermatozoa at room temperature

Reproductive technologies for the generation and maintenance of valuable animal strains

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