Effect of culture time on the spindle and chromosome configurations of human thawed oocytes with slow-cooHng and raid-thawing protocol

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Author:
CONG Lin(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
LI Li(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
ZHANG Zhi-guo(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
WEI Zhao-lian(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
ZHOU Ping(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
ZHAO Ji-hua(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
LI Fen(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
CAO Yun-xia(Reproductive Center,First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)
Journal Title:
CHINESE JOURNAL OF OBSTETRICS AND GYNECOLOGY
Issue:
Volume 43, Issue 02, 2008
DOI:
Key Word:
Oocytes; Cryopreservation; Cells,cultured;Time; Mitotic spindle apparatus;Chromosomes

Abstract: 0bjectlve To examine the metaphase II spindle and chromosome configurations of human oocytes cultured for different times after thawing.Methods Using 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.

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