Vitrification of mouse oocytes using closed pulled straws (CPS) achieves a high survival and preserves good patterns of meiotic spindles, compared with conventional straws, open pulled straws (OPS) and grids.
BACKGROUND
We modified the loading of pulled straws into a new closed system, called closed pulled straws (CPS) for holding oocytes for vitrification. The morphological survival, dynamics of meiotic spindles, and fertilization in vitro of vitrified oocytes using CPS were compared with conventional straws, open pulled straws (OPS), and grids.


METHODS
Surviving oocytes were stained for spindles and chromosomes after 1, 2 and 3 h incubations, and compared with controls. The capacity of fertilization and embryonic cleavage were examined in vitro.


RESULTS
The survival rates of the CPS (79%) and straw (77%) groups were significantly higher (P < 0.05) than the OPS (63%) and grid (39%) groups. At a 1h incubation, vitrified oocytes of four groups had significantly fewer normal spindles than controls (P < 0.05). The straw group was inferior to the others in spindle morphology (P < 0.05). After a 3 h incubation, recovery of vitrified oocytes with normal spindles was significantly improved in all groups (P < 0.05). The percentages of fertilization and blastocyst formation of vitrified oocytes after a 1 h incubation was significantly lower than controls (P < 0.05), but they were improved after 2 or 3 h incubations (P < 0.05).


CONCLUSIONS
Oocytes vitrified using CPS, OPS or grids could lessen spindle injuries and expedite recuperation. The survival using OPS or grids is lower. Sufficient culture time for recovery of meiotic spindle would be imperative for fertilization events of vitrified oocytes. CPS has the advantages of achieving a high survival and preserving good spindles.
Chen
S U
SU
Department of Obstetrics and Gynecology, College of Medicine and The Hospital, National Taiwan University, Taipei, Taiwan.
Lien
Y R
YR
Cheng
Y Y
YY
Chen
H F
HF
Ho
H N
HN
Yang
Y S
YS
D000818:Animals; D002875:Chromosomes / Q000648:ultrastructure; D002970:Cleavage Stage, Ovum; D015925:Cryopreservation / Q000295:instrumentation* / Q000379:methods*; D046508:Culture Techniques; D003599:Cytoskeleton / Q000648:ultrastructure; D005260:Female; D005307:Fertilization in Vitro; D005456:Fluorescent Dyes; D008297:Male; D008540:Meiosis; D051379:Mice; D008813:Mice, Inbred ICR; D008854:Microscopy, Electron; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure*; D012996:Solutions
D005456:Fluorescent Dyes; D012996:Solutions