Influence of cryoprotectant and cooling rate in vitrification method on the spindles of rabbit oocytes
- VernacularTitle:玻璃化冷冻方法中冷冻保护剂及制冷速度对家兔卵母细胞纺锤体的影响
- Author:
Xue-Yong CAI
;
Gui-An CHEN
;
Ying LIAN
;
Xiao-Ying ZHENG
;
Hong-Mei PENG
;
- Publication Type:Journal Article
- Keywords:
Oocytes;
Vitrification;
Cryopreservation;
Spindle;
Rabbits
- From:
Journal of Peking University(Health Sciences)
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.
