Effect of culture time on the spindle and chromosome configurations of human thawed oocytes with slow-cooHng and raid-thawing protocol
- VernacularTitle:人类成熟卵母细胞慢速程序化冷冻复苏后培养时间对纺锤体和染色体的影响
- Author:
Lin CONG
;
Li LI
;
Zhiguo ZHANG
;
Zhaolian WEI
;
Ping ZHOU
;
Jihua ZHAO
;
Fen LI
;
Yunxia CAO
- Publication Type:Journal Article
- Keywords:
Oocytes;
Cryopreservation;
Cells,cultured;
Time;
Mitotic spindle apparatus;
Chromosomes
- From:
Chinese Journal of Obstetrics and Gynecology
2008;43(2):94-97
- CountryChina
- Language:Chinese
-
Abstract:
0bjectlve To examine the metaphase II spindle and chromosome configurations of human oocytes cultured for different times after thawing.MethodsUsing slow.cooling and raid—thawing protocol combined with 0.3 mol/L sucrose and 1.5 mol/L 1,2-propanedio 1(1,2-PROH)to cryoprotect human mature oocytes(n=102),the 64 survival oocytes without abnormal zona pellucida and cytoskeletal were randomly assigned to three groups after thawing:group A:culture 1 hour(n=20),group B:culture 3 hour(n=22),group C:culture 5 hours(n=22),the flesh oocytes served as control group(n=18).Immunocytochefifical staining and fluorescence microscopy were used to assess the morphology of the metaphase II spindle and chromosome.Results(1)The normal spindle rates of groups A,B and C were 10％(2/20),46％(10/22)and 41％(9/22)respectively,significantly decreased compared with control group(83％,15/18;P<0.05).The rates of absent spindle in group A(45％,9/20)was significantly higher than control group(6％,1/18;P<0.01).Also,the rates of absent spindle in group A was higher than groups B(14％,3/20)and C(14％,3/20;P<0.05).However,no significant differences were observed in groups B and C(P>0.05).(2)A significant increase in abnormal chromosome rate was observed in group A(30％,6/20)compared to groups B(68％,15/22),C(64％,14/22)and control group(78％,14/18;尸<0.05).No differences in chromosome morphology were observed in groups B,C and control group(P>0.05).Conclusions The cryoproteetant protocol leads to a deleterious effect on the organization of the meiotic spindle and chromosome at MI stage.The 3—5 hours post—thawing incubation could permit restoration of the meiotic spindles and chromosome.
