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Published on: December 12, 2011
Transportation of freeze-dried mouse spermatozoa under different preservation conditions
Yosuke Kawase1, Takanori Tachibe, Kou-ichi Jishage
1Chugai Research Institute for Medical Science, Shizuoka, Japan.
Abstract:
Freeze-dried mouse spermatozoa can be used for normal embryonic development after injection into oocytes, thus indicating that freeze-drying is a useful method for the storage and transportation of genetic materials from animals. We recently reported that storage of freeze-dried mouse spermatozoa requires maintenance at temperatures lower than -80 C for long-term preservation and a pressure of 0.37 mbar at primary drying and that these conditions significantly improve the developmental rate to the blastocyst stage. In this study, we examined the influence of transportation and preservation conditions on freeze-dried spermatozoa. Freeze-dried spermatozoa stored for 2 or 2.5 years at 4 or -80 C were transported round trip overland between Shizuoka and Hokkaido prefectures in Japan or by air between Japan and Belgium. The freeze-drying conditions consisted of primary drying at pressures of 0.04, 0.37 and 1.03 mbar and secondary drying at a pressure of 0.001 mbar. Embryos (2-cell stage) from freeze-dried spermatozoa dried at 0.04 mbar and stored at 4 C for 2 years with and without overland transportation did not develop to term. The development rates of embryos from spermatozoa stored at -80 C for up to 2 years and transported overland, by air and without transportation were 8, 1 and 28%, respectively. The development rates of embryos from spermatozoa without transportation were significantly higher than with transportation (P<0.05). These data indicate that freeze-dried spermatozoa stored at -80 C with and without transportation can retain their ability to generate viable offspring after storage for up to 2 years. However, there are limitations to be considered in the transportation of freeze-dried spermatozoa at ambient temperature.
Insights
Freeze-dried mouse spermatozoa can be stored for up to two years at -80°C and still produce viable offspring, even after transportation. However, ambient temperature transport significantly reduces their developmental potential.
Area of Science:
- Reproductive biology
- Cryopreservation
- Animal genetics
Background:
- Freeze-drying (lyophilization) is a viable method for preserving animal genetic material.
- Previous research indicated optimal long-term storage conditions for freeze-dried mouse spermatozoa involve temperatures below -80°C and specific drying pressures.
Purpose of the Study:
- To investigate the impact of various transportation and preservation conditions on the viability of freeze-dried mouse spermatozoa.
- To determine the long-term efficacy of freeze-dried spermatozoa after simulated transport.
Main Methods:
- Freeze-dried mouse spermatozoa were stored for 2-2.5 years at either 4°C or -80°C.
- Samples underwent simulated transportation via overland (Japan) or air (Japan-Belgium) routes.
- Developmental rates to term were assessed after intracytoplasmic sperm injection into oocytes.
Main Results:
- Spermatozoa stored at 4°C, regardless of transportation, failed to produce viable offspring.
- Spermatozoa stored at -80°C without transportation showed the highest development rate (28%).
- Transport, both overland (8%) and by air (1%), significantly reduced the developmental potential of spermatozoa stored at -80°C compared to no transport (P<0.05).
Conclusions:
- Freeze-dried mouse spermatozoa stored at -80°C maintain their ability to generate viable offspring for up to two years, even with transportation.
- Transportation at ambient temperatures poses a significant challenge to the viability of freeze-dried spermatozoa.
- Optimal preservation and transport protocols are crucial for utilizing freeze-dried spermatozoa for genetic material preservation.
