Effect of embryo density and microdrop volume on the blastocyst development of mouse two-cell embryos.
OBJECTIVE
To study the effect of embryo density and microdrop volume on mouse two-cell embryo development.


METHODS
Experimental study.


METHODS
Assisted conception laboratory.


METHODS
Two-cell mouse embryos (n = 1511).


METHODS
One, five, 10, or 15 embryos were cultured in 10-μL drops of cleavage medium. In the second study, embryos were cultured singly in 0.5-, 1-, 2-, 5-, and 10-μL drops. Finally, they were cultured in pair in 0.5-, 1-, and 2-μL drops. After 48 hours, embryos were transferred into blastocyst medium for an additional 24 hours.


METHODS
Cleavage and blastocyst formation and inner cell mass (ICM) and trophectoderm (TE) cell numbers.


RESULTS
No differences in cleavage or blastocyst formation were found in different groups in experiment 1, 2, or 3. Embryos cultured singly had fewer ICM and TE cells than those cultured in groups. Embryos cultured singly in 0.5 μL had fewer TE cells than those in 10 μL, but had insignificant difference in the ICM. Duo culture in 0.5-2 μL appeared to give the same results as group culture in 10-μL drops.


CONCLUSIONS
Group culture is preferred when using sequential media. Beneficial effects cannot be mimicked by volume reduction in single-embryo culture.
Vutyavanich
Teraporn
T
Division of Reproductive Medicine, Department of Obstetrics and Gynecology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand. tvutyava@med.cmu.ac.th
Saeng-Anan
Ubol
U
Sirisukkasem
Suppachai
S
Piromlertamorn
Waraporn
W
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D002452:Cell Count / Q000379:methods; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR
D003470:Culture Media