|
Objective: To investigate the use vitrification freezing of ovarian tissue preservation. Find frozen ovarian tissue is more appropriate for the protection of liquid formulations. Methods: collection a 10-14dpc ICR mice ovaries and 2 cases of human ovarian tissue samples (cut into approximately 1 × 1 × 1-2mm3 tissue blocks), using four different vitrification protective solution: 40% ethylene glycol 18 % Ficoll-70 0.3 M sucrose; 40% propylene glycol 18% Ficoll-70 0.3 M sucrose; 2.62M 1.31M dimethyl sulfoxide, propylene glycol 0.0075 polyethylene glycol 2.6M acetamide; 1.5M Propylene glycol 0.1 M sucrose ovarian tissue -205 ° C surface vitrification. By morphological evaluation of follicle activity in mice and human ovarian tissue before and after the Thaw, the effect of cryopreservation for testing. Results: The mouse ovarian tissue vitrification experiments to compare the the four glass transition cryoprotectant cryopreservation of mouse ovarian preantral follicles recovery rate in vitro and adherence rates were significant between any two The difference (P lt; 0.01). In which the cryoprotectant A recovery rate and 1 day, 5 days follicles adherence rates were 55.13%, 30.90% and 27.05%, standard deviation (SD) were 4.72,1.60 and 4.28 respectively, the coefficient of variation (CV) 8.56,5.18 and 15.82; the cryoprotectant B recovery rate and 1 day, 5 days follicle adherence rates were 42.74%, 25.05% and 18.47%, respectively 3.30,5.27 and 5.02 standard deviation (SD), coefficient of variation (CV) were 7.72,21.02 and 27.19; cryoprotectant C recovery rate and 1 day, 5 days adherent rates were 62.79%, 39.39% and 32.68%, standard deviation (SD) were 5.83,2.37 and 3.13 the coefficient of variation (CV) were 9.28,6.02 and 9.59; cryoprotectant D recovery rate and 1 day, 5 days adherence rates were 23.89%, 13.31% and 7.87%, standard deviation (SD) were 3.90, 3.67 and 2.50, coefficient of variation (CV) were 16.31,27.55 and 31.79. Cryoprotectant C cryopreservation best results. In human ovarian tissue vitrification experiment, cryoprotectant group there are significant differences (P lt; 0.005) and normal tissue. The best frozen protective effect of cryo-protectant solution C (P lt; 0.005), after the recovery of the frozen tissue 63.41% of morphologically normal follicles follicles in fresh tissue group morphologically normal rate of 89.86%. No significant difference in results between the cryoprotective solution A, B and D groups (P GT; 0.05) (30.26%, 36.23% and 23.60%). Conclusion: 1.-205 ° C surface vitrification is a simple and effective ovarian freezing methods. 2 the cryoprotectant C is a good preservation of ovarian tissue vitrification protective solution.
|