In vitro blastocyst development from serially split mouse embryos and future implications for human assisted reproductive technologies.
OBJECTIVE
To assess the efficacy of serial splitting of mouse embryos with respect to blastocyst development.


METHODS
Prospective study.


METHODS
Commercial research facility.


METHODS
Commercially available mouse embryos from B6C3F-1 x B6D2F-1.


METHODS
One, two, and three blastomeres were biopsied from two-, four-, and six-cell embryos, respectively, and were inserted into empty zona pellucida recipients (first split). These embryos were cultured to reach their original cell number status and then were split again (second split). Once these embryos regained their original cell status, they were split yet again (third split).


METHODS
Blastocyst development of embryos split serially at the two-, four-, and six-cell stages.


RESULTS
The blastocyst development rate for two-, four-, and six-cell embryos subjected to a first split was 74.3%, 75.0%, and 66.6%, respectively, as compared with 71.8%, 62.6%, and 48.4% (second split) and 48.4%, 38.1%, and 10.6% (third split).


CONCLUSIONS
First and second splitting of cleavage-stage embryos has yielded high efficiency rates for blastocyst development when compared with the third splitting, which did not provide any beneficial advantage for further embryo splitting and multiplication. This is the first study reporting on three serial embryo splittings in a mammalian species. Embryo splitting may have significant impact and applications in human assisted reproductive technology.
Illmensee
Karl
K
Reprogen Ltd., Limassol, Cyprus.
Kaskar
Khalied
K
Zavos
Panayiotis M
PM
D000818:Animals; D001706:Biopsy / Q000379:methods*; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D004624:Embryo Transfer; D047108:Embryonic Development; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D008866:Microsurgery; D027724:Reproductive Techniques, Assisted