Preliminary Study of Human Ovarian Tissue Cryopreservation by Controlled-rate Freezing
- VernacularTitle:人卵巢组织慢速程序化冷冻保存方案的初步探讨
- Author:
Liying LIU
;
Wenyu QU
;
Li JIANG
;
Xiaoli ZHANG
;
Xiaoli LIU
- Publication Type:Journal Article
- Keywords:
cryopreservation;
human ovarian tissues;
ultrastructure;
controlled-rate freezing
- From:
Journal of China Medical University
2015;(5):425-428,433
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the effect of different cryoprotectants and different concentrations on controlled?rate freezing of human ovarian tissues. Methods Ovarian tissues were sampled from 15 patients undergoing benign ovarian tumor surgery. Cortical slices were frozen by controlled?rate freezing using three cryoprotectants,propanediol,ethanediol,and dimethylsulphoxide,and the concentration of each cryoprotectant was 1.5 mol/L or 2.0 mol/L. Cortical slices obtained from each patient were processed with each cryopreservation procedure simultaneously. Morphology of fol?licles was studied by light and electron microscopy and the normal rate was compared with that of the fresh tissues from the patient. Results There were no significant differences in distribution of follicles of different developmental stages between each group(P<0.05). Light microscopy showed 1.5 mol/L EG and 2.0 mol/L EG groups had the best freezing effect,and the difference in the morphologically normal rate of follicles was not statisti?cally significant compared to fresh controls(P>0.05). However,the difference was statistically significant for 1.5 mol/L PROH,2.0 mol/L PROH, 1.5 mol/L DMSO and 2.0 mol/L DMSO groups(P<0.05). Electron microscopy showed the oocyte quality declined after controlled?rate freezing pro?cedure. However,the statistical analysis was not conducted due to little data. The viability of granulosa cells was significantly declined after all the freezing procedures compared to that of the fresh control tissues(P<0.01). The number of morphologically normal granulosa cells was slightly higher in the tissues which had been cryopreserved with 2.0 mol/L PROH and 1.5 mol/L EG,but no significant differences were found between any of two frozen groups(P>0.05). Conclusion Controlled?rate freezing using 1.5 mol/L EG as the cryoprotectant can better save oocytes and granulosa cells. It is a preferable freezing procedure for ovarian tissues.
