人卵巢组织冻融过程对卵巢组织的影响

doi:10.3969/j.issn.1006-7795.2025.04.002

首都医科大学学报 ›› 2025, Vol. 46 ›› Issue (4): 594-600.doi: 10.3969/j.issn.1006-7795.2025.04.002

• 更年期妇科内分泌与生育力保护 • 上一篇下一篇

人卵巢组织冻融过程对卵巢组织的影响

李扬璐¹,阮祥燕^1*,杜娟¹,程姣姣¹,金凤羽¹,谷牧青¹,李妍秋¹,Alfred O. Mueck^1,2

1.首都医科大学附属北京妇产医院/北京妇幼保健院内分泌科，北京 100026；2.德国图宾根大学妇产医院妇女健康部与妇女健康研究中心，图宾根 D-72076

收稿日期:2024-12-30 出版日期:2025-08-21 发布日期:2025-08-29
通讯作者: 阮祥燕 E-mail:ruanxiangyan@ccmu.edu.cn
基金资助:
国家自然科学基金项目（82101822），北京市属医院科研培育计划项目（PX2021053），首都医科大学附属北京妇产医院中青年学科骨干培养专项（FCYY202015），首都医科大学校培育基金（PYZ20058），北京市自然科学基金项目（7202047）。

Impact of cryopreservation and thawing on human ovarian tissue

Li Yanglu¹, Ruan Xiangyan^1*, Du Juan¹, Cheng Jiaojiao¹, Jin Fengyu¹, Gu Muqing¹, Li Yanqiu¹, Alfred O. Mueck^1,2

1.Department of Gynecological Endocrinology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Health Care Hospital, Beijing 100026, China ；2.Department of Women's Health, Research Centre for Women's Health and University Women's Hospital of Tuebingen, University Hospitals of Tuebingen, Tuebingen D-72076, Germany

Received:2024-12-30 Online:2025-08-21 Published:2025-08-29
Supported by:
This study was supported by National Natural Science Foundation of China(82101822)， Beijing Municipal Administration of Hospitals Incubating Program (PX2021053)， Beijing Obstetrics and Gynecology Hospital, Capital Medical University (FCYY202015)， Capital Medical University (PYZ20058)， Natural Science Foundation of Beijing (7202047).

摘要/Abstract

摘要： 目的采用慢速冷冻方法，以首都医科大学附属北京妇产医院生育力保护中心标准冻融方案，评估人卵巢组织冻融过程对卵巢组织的影响。方法将12例患者的卵巢组织分为新鲜卵巢组织组和冻融卵巢组织组，冻融方案为本中心标准方案，通过苏木精-伊红 ( hematoxylin-eosin，HE)染色法和钙黄绿素AM（calcein acetoxymethylester，calcein-AM）染色进行卵泡计数并评估卵泡活性，免疫组织化学法标记Ki-67和caspase-3的阳性率，评估细胞增殖和凋亡情况，Western blotting法比较两组在caspase-3、bax、FasL等凋亡相关蛋白的表达。结果采用本中心标准冻融方案，卵巢组织冻融前后在卵泡计数和卵泡活性方面差异无统计学意义（P>0.05）；卵巢组织冻融后细胞Ki-67阳性率低于新鲜卵巢组织（P<0.05），差异有统计学意义；两组caspase-3阳性率差异无统计学意义（P>0.05）；冻融后caspase-3蛋白表达量高于新鲜卵巢组织，差异有统计学意义（P<0.05）；而bax和FasL等凋亡相关蛋白的表达差异无统计学意义（P>0.05）。结论采用本中心标准冻融方案，能够很好地保持卵巢组织内的卵泡数目、形态和活性，冻融后细胞增殖水平降低，不增加冻融后bax和FasL等凋亡相关蛋白的表达，caspase-3的表达量相对增加，是较好的人卵巢组织冻融方案。

关键词: 卵巢组织, 冻融方案, 卵泡活性, 凋亡相关蛋白, 生育力保护, 卵泡损伤

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

中图分类号:

李扬璐, 阮祥燕, 杜娟, 程姣姣, 金凤羽, 谷牧青, 李妍秋, Alfred O. Mueck. 人卵巢组织冻融过程对卵巢组织的影响[J]. 首都医科大学学报, 2025, 46(4): 594-600.

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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人卵巢组织冻融过程对卵巢组织的影响

Impact of cryopreservation and thawing on human ovarian tissue

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