Vitrification of mouse oocytes using a nylon loop.
Cryopreservation of mouse oocytes was improved by the use of ultra-rapid vitrification using a nylon loop of 0.5 mm diameter. Oocytes that were vitrified using the loop survived at high rates and were fertilized following a small hole being made in the zona pellucida (69.8%) and developed to the blastocyst stage in culture (67.4%) at similar rates to that of oocytes that were not cryopreserved. Blastocysts resulting from oocytes vitrified using the nylon loop had similar development of the inner cell mass and trophectoderm as blastocysts from non-cryopreserved oocytes. In contrast, oocytes that were cryopreserved using a slow-freezing protocol where most of the Na+ is replaced with choline had lower rates of fertilization (39.5%), reduced development to the blastocyst stage (25.7%), and blastocysts had reduced development of the inner cell mass. Blastocysts derived from oocytes that were vitrified with the nylon loop were able to implant (88.0%) and develop into fetuses (56.5%) at significantly higher rates compared to blastocysts derived from oocytes that were slow-frozen (52.4 and 26.2%, respectively). Vitrification of mouse oocytes using the nylon loop results in the retention of viability of the oocytes and subsequent embryos.
Lane
M
M
Colorado Center for Reproductive Medicine, Research and Development, Englewood, Colorado 80110, USA. mlane@colocrm.com
Gardner
D K
DK
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D046508:Culture Techniques; D004155:Diphenhydramine / Q000031:analogs & derivatives; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000502:physiology*; D011897:Random Allocation
D004155:Diphenhydramine; C005715:embramine