The oocyte spindle is preserved by 1,2-propanediol during slow freezing.
OBJECTIVE
To investigate the specific changes in oocyte spindle subjected to severe challenges of low temperature, as well as to examine the effect of cryoprotectants in preserving oocyte spindle during cryopreservation.


METHODS
In vitro experimental study.


METHODS
Academic research laboratory.


METHODS
B6D2F1 (C57BL/6 X DBA/2) mice.


METHODS
Mouse oocytes were cryopreserved using a slow freezing method in a sodium-depleted medium with 1.5 mol/l 1,2-propanediol (PROH) and 0.3 M sucrose. To examine the spindle, oocytes were fixed before, during, and after cryopreservation, and oocytes were analyzed by immunocytochemistry and confocal microscopy.


RESULTS
The MII spindle was preserved during the slow freezing, because the cryoprotectant PROH was found to support the organization of MII spindle in resisting the subzero temperature. In contrast, the MII spindle was disassembled gradually during the thawing process with or without PROH. Most of the oocytes were able to recover the MII spindle after thawing, but a portion of thawed oocytes could not sustain the meiotic spindle because of parthenogenetic activation.


CONCLUSIONS
1,2-Propanediol can support the organization of MII spindle to defy the subphysiologic temperature; however, the PROH cannot sustain oocyte spindle structure after the subsequent thawing process.
Chang
Ching-Chien
CC
Reproductive Biology Associates, 1150 Lake Hearn Drive, Suite 600, Atlanta, GA 30342, USA. peter.nagy@rba-online.com
Sung
Li-Ying
LY
Lin
Chih-Jen
CJ
Kort
Hilton I
HI
Yang
Xiangzhong
X
Tian
X Cindy
XC
Nagy
Zsolt Peter
ZP