Successful long-term preservation of rat sperm by freeze-drying |
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Authors: | Kaneko Takehito Serikawa Tadao |
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Institution: | Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto, Japan. tkaneko@anim.med.kyoto-u.ac.jp |
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Abstract: | BackgroundFreeze-drying sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying sperm is that it can be stored at 4°C and transported at room temperature. Although the successful freeze-drying of sperm has been reported in a number of animals, the possibility of long-term preservation using this method has not yet been studied.Methodology/Principal FindingsOffspring were obtained from oocytes fertilized with rat epididymal sperm freeze-dried using a solution containing 10 mM Tris and 1 mM EDTA adjusted to pH 8.0. Tolerance of testicular sperm to freeze-drying was increased by pre-treatment with diamide. Offspring with normal fertility were obtained from oocytes fertilized with freeze-dried epididymal sperm stored at 4°C for 5 years.Conclusions and SignificanceSperm with –SS– cross-linking in the thiol-disulfide of their protamine were highly tolerant to freeze-drying, and the fertility of freeze-dried sperm was maintained for 5 years without deterioration. This is the first report to demonstrate the successful freeze-drying of sperm using a new and simple method for long-term preservation. |
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