Evaluation of developmental competence of vitrified-warmed early cleavage stage embryos and their application for chimeric mouse production.
The developmental competence of in vitro cultured embryos vitrified-warmed at an early cleavage stage (2- or 4, 8-cell stage) was examined by both direct transfer into recipient animals and after in vitro manipulation for chimeric mice production using embryonic stem (ES) cells. Vitrified-warmed embryos transferred at the morulae and blastocyst stages showed fetus development comparable to control embryos, although blastocyst development of vitrified-warmed embryos was significantly slower than that of controls. When vitrified-warmed early cleavage stage embryos were used for chimeric mouse production using ES cells, 1 to 10% of the injected or aggregated embryos developed into chimeric neonates and germ-line chimeric mice were obtained from all ES cell lines. This study indicates that embryos developed in vitro from vitrified-warmed embryos have equivalent competence with unvitrified embryos irrespective of stage of vitrification and that these vitrified-warmed embryos maintain adequate viability even after in vitro manipulation such as aggregation and microinjection with ES cells.
Kito
Seiji
S
National Institute of Radiological Sciences, 4-6-1 Anagawa, Chiba 263-8555, Japan.
Noguchi
Yoshiko
Y
Ohta
Yuki
Y
Ohhata
Tatsuya
T
Abe
Masumi
M
Shiomi
Naoko
N
Shiomi
Tadahiro
T
D000818:Animals; D002678:Chimera; D004622:Embryo, Mammalian / Q000166:cytology*; D051379:Mice