High survival of rabbit morulae after vitrification in an ethylene glycol-based solution by a simple method.
Rabbit morulae were exposed to a vitrification solution-modified PBS [PB1] medium containing 40% ethylene glycol + 18% Ficoll + 0.3 M sucrose (EFS) for 2, 5, or 10 min at 20 degrees C and were vitrified in liquid nitrogen. When morulae were rapidly warmed, 96% had an intact zona pellucida. When embryos were cultured after removal of the mucin coat, high proportions of them formed blastocoel (79-100%), but the percentage of embryos developed to fully expanded blastocysts decreased with increased exposure time 87%, 40%, and 17%). The survival rate of morulae vitrified after removal of the mucin coat was lower than that of mucin-intact embryos. To assess the development potential in vivo, 131 embryos were vitrified after 2 min of exposure to EFS solution; all the embryos were recovered and 120 were transferred to recipients without removal of the mucin coat, resulting in 78 (65%) full-term fetuses or young. This simple method, which yields high survival both in vitro and in vivo, will be of practical use for vitrifying rabbit embryos.
Kasai
M
M
Laboratory of Animal Science, College of Agriculture, Kochi University, Japan.
Hamaguchi
Y
Y
Zhu
S E
SE
Miyake
T
T
Sakurai
T
T
Machida
T
T
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D046508:Culture Techniques; D004624:Embryo Transfer; D019855:Ethylene Glycol; D005026:Ethylene Glycols; D005260:Female; D009028:Morula / Q000502:physiology*; D009077:Mucins / Q000502:physiology; D009584:Nitrogen; D011817:Rabbits / Q000196:embryology*; D012996:Solutions; D015044:Zona Pellucida / Q000502:physiology
D005026:Ethylene Glycols; D009077:Mucins; D012996:Solutions; D019855:Ethylene Glycol; D009584:Nitrogen