title: [Effect of culture time on the spindle and chromosome configurations of human thawed oocytes with slow-cooling and raid-thawing protocol].
abstract: OBJECTIVE
  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 fresh oocytes served as control group (n = 18). Immunocytochemical 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%, 0; 6/20) compared to groups B (68%, 15/22), C (64%,14/22) and control group (78%, 14/18; P < 0.05). No differences in chromosome morphology were observed in groups B, C and control group (P > 0.05).
  
  
  CONCLUSIONS
  The cryoprotectant protocol leads to a deleterious effect on the organization of the meiotic spindle and chromosome at MII stage. The 3 - 5 hours post-thawing incubation could permit restoration of the meiotic spindles and chromosome.
authors.affiliation: Reproductive Center, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Cong
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Zhi-Guo
authors.initials: ZG
authors.lastname: Zhang
authors.forename: Zhao-Lian
authors.initials: ZL
authors.lastname: Wei
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.forename: Ji-Hua
authors.initials: JH
authors.lastname: Zhao
authors.forename: Fen
authors.initials: F
authors.lastname: Li
authors.forename: Yun-Xia
authors.initials: YX
authors.lastname: Cao
mesh_terms: D002470:Cell Survival; D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000648:ultrastructure*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D005260:Female; D006801:Humans; D008540:Meiosis; D008677:Metaphase; D018613:Microscopy, Confocal; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure*; D008941:Spindle Apparatus / Q000648:ultrastructure*; D013997:Time Factors
chemical_list: D003451:Cryoprotective Agents