15 research outputs found
Transportation of Freeze-Dried Mouse Spermatozoa Under Different Preservation Conditions
Transportation of Freeze-Dried Mouse Spermatozoa Under Different Preservation Conditions
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.http://www.jstage.jst.go.jp/article/jrd/53/6/53_1169/_article/-char/ja
Effect of Zona Incision by Piezo-Micromanipulator (ZIP) on In Vitro Fertilization in 21 Transgenic Mice Lines
Effect of Zona Incision by Piezo-Micromanipulator (ZIP) on In Vitro Fertilization in 21 Transgenic Mice Lines
Zona incision using a piezo-micromanipulator (ZIP) has been demonstrated to be effective for in vitro fertilization (IVF) using cryopreserved C57BL/6 spermatozoa. In this study, ZIP oocytes inseminated with frozen-thawed genetically modified C57BL/6J or FVB mice spermatozoa (21 lines) showed fertilization rates of 22-75% and live fetus rates of 8-49%. In 6 of the lines, the fertilization rates for oocytes compared with ZIP (42-75%) were significantly higher than that of nontreated oocytes (0-50%). Using only 90 oocytes for IVF with ZIP, 5 breeding pairs were produced from cryopreserved genetically modified mice spermatozoa. Our results indicate that application of the ZIP technique is effective for IVF using cryopreserved genetically modified mouse spermatozoa.http://www.jstage.jst.go.jp/article/expanim/58/4/58_415/_article/-char/ja