Impact of cryopreservation and thawing on human ovarian tissue

doi:10.3969/j.issn.1006-7795.2025.04.002

Abstract

Abstract: Objective To evaluate the impact of slow-freezing process on human ovarian tissue with the standard cryopreservation-thawing protocol of Fertility Protection Center of Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Methods Ovarian tissues of 12 patients were divided into fresh ovarian tissue group （fresh group）and freezing-thawing ovarian tissue group (F-T group). The freezing-thawing protocol was the standard protocol in our center. The number and activity of follicle were examined with Hematoxylin-eosin (HE) staining and calcein-AM (calcein acetoxymethylester) staining, and the proliferation and apoptosis was evaluated with the immunohistochemical staining of Ki-67 and caspase-3. The expressions of apoptosis-related proteins such as caspase-3, bax and FasL between the two groups were compared with Western blotting. Results There were no statistically significant differences in follicle counting and follicle activity in ovarian tissues pre- and post- freezing-thawing (P>0.05), and the positive rate of Ki-67 in ovarian tissues after freezing-thawing was significantly lower than that in fresh ovarian tissues (P<0.05), and there was no statistically significant difference in the positive rate of caspase-3 between the two groups (P>0.05). The expression of caspase-3 protein in ovarian tissues after freezing-thawing was significantly higher than that in fresh ovarian tissues (P<0.05), while the expressions of other apoptosis-related proteins such as bax and FasL were not significantly different (P>0.05). Conclusion The standard cryopreservation-thawing regimen in our center can effectively maintain the follicle number, morphology, and activity in ovarian tissues. After freezing and thawing, the cell proliferation level is decreased. The expression of apoptosis-related proteins such as bax and FasL are not increased, and the expression of caspase-3 is relatively increased. These results suggest our freezing-thawing regimen is good for human ovarian tissue.

Key words: ovarian tissue, freezing and thawing protocol, follicle viability, apoptosis-related protein, fertility protection, follicle injury

CLC Number:

Li Yanglu, Ruan Xiangyan, Du Juan, Cheng Jiaojiao, Jin Fengyu, Gu Muqing, Li Yanqiu, Alfred O. Mueck. Impact of cryopreservation and thawing on human ovarian tissue[J]. Journal of Capital Medical University, 2025, 46(4): 594-600.

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