天然冷冻保护剂L-脯氨酸与传统冷冻保护剂冻存的人卵巢组织移植有效性研究

doi:10.3969/j.issn.1006-7795.2026.04.006

首都医科大学学报 ›› 2026, Vol. 47 ›› Issue (4): 673-679.doi: 10.3969/j.issn.1006-7795.2026.04.006

• 更年期妇科内分泌与生育力保护 • 上一篇下一篇

天然冷冻保护剂L-脯氨酸与传统冷冻保护剂冻存的人卵巢组织移植有效性研究

柳岸鸣¹,阮祥燕^1*,杜娟¹,蒋玲玲¹,李妍秋¹,谷牧青¹,程姣姣¹, 张明珍¹,王泽铖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:2026-04-09 修回日期:2026-05-05 出版日期:2026-08-21 发布日期:2026-07-26
通讯作者: 阮祥燕 E-mail:ruanxiangyan@ccmu.edu.cn
基金资助:
北京市医院管理局“登峰”人才培养计划项目(DFL20181401)，北京市卫生健康委员会北京市研究型病房示范建设项目(BCRW202109)，国家外国专家个人类项目生育力保护与妇科内分泌疑难病研究会诊中心的建立 (S20240236)，国家外国专家个人类项目全国女性卵巢保护与抗衰促进工程项目建设(S20240230) 。

Efficacy of the natural cryoprotectant L-proline versus conventional cryoprotectants in human ovarian tissue cryopreservation and transplantation

Liu Anming¹, Ruan Xiangyan^1*, Du Juan¹, Jiang Lingling¹, Li Yanqiu¹, Gu Muqing¹, Cheng Jiaojiao¹, Zhang Mingzhen¹, Wang Zecheng¹, 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:2026-04-09 Revised:2026-05-05 Online:2026-08-21 Published:2026-07-26
Supported by:
This study was supported by Beijing Municipal Administration of Hospitals’ Ascent Plan (DFL20181401), Beijing Municipal Health Commission, Demonstration Construction Project of Clinical Research Ward (BCRW202109), National Foreign Experts Individual Project, Establishment of Fertility Preservation and Consultation Center for Gynecological Endocrine Difficult Diseases (S20240236), National Foreign Experts Individual Project, Construction of National Female Ovarian Preservation and Anti-aging Promotion Project (S20240230).

摘要/Abstract

摘要： 目的探索天然冷冻保护剂L-脯氨酸（L-proline, L-pro）冷冻保存人卵巢组织移植的有效性，并与传统冷冻保护剂二甲基亚砜（dimethyl sulfoxide，DMSO）进行比较。方法采用慢速冷冻方法，将10例患者的卵巢皮质组织分别用基于DMSO和L-pro的冻存液进行冷冻保存，复苏后移植于已去势雌性裸鼠体内；对照组只去势。移植21 d后取出卵巢组织移植物进行卵泡计数，并检测血清中促卵泡刺激素（follicle-stimulating hormone, FSH），雌二醇（estradiol, E2）和抗米勒管激素（anti-Müllerian Hormone, AMH）的浓度。结果 L-pro组静止卵泡数量显著高于DMSO组（P=0.042），而2组在生长卵泡数量及总卵泡数量方面差异无统计学意义（P均>0.05）。2个实验组各项血清激素水平较对照组均显著改善（P均<0.05），且L-pro组与DMSO组间各项血清激素水平差异无统计学意义。结论 L-pro作为冷冻保护剂在卵巢组织冻存异种移植中卵泡保护和激素恢复的效果不亚于传统的冷冻保护剂DMSO。为卵巢组织冻存的冷冻保护剂优化提供了新的思路和实验依据，未来仍需进一步深化机制研究和长期功能的研究。

关键词: L-脯氨酸, 天然冷冻保护剂, 卵巢组织冻存, 异种移植, 生育力保存, 卵泡

Abstract: Objective To evaluate the efficacy of the natural cryoprotectant L-proline (L-pro) in human ovarian tissue cryopreservation and transplantation, and to compare it with the conventional cryoprotectant dimethyl sulfoxide (DMSO).Methods Ovarian cortical tissues obtained from ten patients were cryopreserved using a slow-freezing protocol with either DMSO-based or L-pro-based cryoprotective solutions. After thawing, the tissues were xenotransplanted into ovariectomized female nude mice, while the control group underwent ovariectomy only. Twenty-one days after transplantation, grafts were retrieved for follicle counting, and serum levels of follicle-stimulating hormone (FSH), estradiol (E2), and anti-Müllerian hormone (AMH) were measured.Results The number of quiescent (primordial) follicles was significantly higher in the L-pro group than in the DMSO group (P=0.042), whereas no significant differences were observed in the number of growing follicles or total follicle counts between the two groups (all P> 0.05). Serum hormone levels in both experimental groups were significantly improved compared with the control group (all P<0.05), with no significant differences between the L-pro and DMSO groups.Conclusion L-pro, as a cryoprotectant, demonstrates comparable efficacy to DMSO in preserving follicular integrity and restoring endocrine function following ovarian tissue cryopreservation and xenotransplantation. These findings provide a novel perspective for optimizing cryoprotective agents in ovarian tissue preservation, although further studies are needed to elucidate the underlying mechanisms and long-term functional outcomes.

Key words: L-proline, natural cryoprotectant, ovarian tissue cryopreservation, xenotransplantation, fertility preservation, follicle

中图分类号:

柳岸鸣, 阮祥燕, 杜娟, 蒋玲玲, 李妍秋, 谷牧青, 程姣姣, 张明珍, 王泽铖, Alfred O. Mueck. 天然冷冻保护剂L-脯氨酸与传统冷冻保护剂冻存的人卵巢组织移植有效性研究[J]. 首都医科大学学报, 2026, 47(4): 673-679.

Liu Anming, Ruan Xiangyan, Du Juan, Jiang Lingling, Li Yanqiu, Gu Muqing, Cheng Jiaojiao, Zhang Mingzhen, Wang Zecheng, Alfred O. Mueck, . Efficacy of the natural cryoprotectant L-proline versus conventional cryoprotectants in human ovarian tissue cryopreservation and transplantation[J]. Journal of Capital Medical University, 2026, 47(4): 673-679.

参考文献

［1］Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA Cancer J Clin, 2024, 74(3): 229-263.
［2］Vidal A, Bora C, Jarisch A, et al. Impact of haematopoietic stem cell transplantation for benign and malignant haematologic and non-haematologic disorders on fertility: a systematic review and meta-analysis［J］. Bone Marrow Transplant, 2025, 60(5): 645-672.
［3］Ruan X Y, Xu C, Huang H F, et al. Practice guideline on ovarian tissue cryopreservation and transplantation in the prevention and treatment of iatrogenic premature ovarian insufficiency［J］. Maturitas, 2024, 182: 107922.
［4］Rebar R W, Keator C S. Expanding our knowledge of premature ovarian insufficiency［J］. Fertil Steril, 2021, 115(2): 328-329.
［5］Lambertini M, Peccatori F A, Demeestere I, et al. Fertility preservation and post-treatment pregnancies in post-pubertal cancer patients: ESMO Clinical Practice Guidelines†［J］. Ann Oncol, 2020, 31(12): 1664-1678.
［6］Su H I, Lacchetti C, Letourneau J, et al. Fertility preservation in people with cancer: ASCO guideline update［J］. J Clin Oncol, 2025, 43(12): 1488-1515.
［7］Khattak H, Malhas R, Craciunas L, et al. Fresh and cryopreserved ovarian tissue transplantation for preserving reproductive and endocrine function: a systematic review and individual patient data meta-analysis［J］. Hum Reprod Update, 2022, 28(3): 400-416.
［8］Ruan X Y, Du J, Jin F Y, et al. Second pregnancy in China 5 years after ovarian tissue transplantation to prevent iatrogenic POI［J］. Climacteric, 2025, 28(2): 212-216.
［9］Ruan X, Cheng J, Korell M, et al. Ovarian tissue cryopreservation and transplantation prevents iatrogenic premature ovarian insufficiency: first 10 cases in China［J］. Climacteric, 2020, 23(6): 574-580.
［10］Lewis J K, Bischof J C, Braslavsky I, et al. The grand challenges of organ banking: proceedings from the first global summit on complex tissue cryopreservation［J］. Cryobiology, 2016, 72(2): 169-182.
［11］Mazur P. Freezing of living cells: mechanisms and implications［J］. Am J Physiol, 1984, 247(3 Part 1): C125-C142.
［12］Best B P. Cryoprotectant toxicity: facts, issues, and questions［J］. Rejuvenation Res, 2015, 18(5): 422-436.
［13］Ding Y Q, Liu S, Liu J T, et al. Cryopreservation with DMSO affects the DNA integrity, apoptosis, cell cycle and function of human bone mesenchymal stem cells［J］. Cryobiology, 2024, 114: 104847.
［14］Chen X, Zhan T J, Wang Y T, et al. Dimethyl-sulfoxide-free cell cryoprotectant derived from amino acids［J］. Langmuir, 2024, 40(49): 25919-25930.
［15］Abdelnour S A, Khalil W A, Khalifa N E, et al. L-Proline: a promising tool for boosting cryotolerance and fertilizing ability of cryopreserved sperm in animals［J］. Anim Reprod Sci, 2024, 263: 107429.
［16］Qin Q Y, Zhao L S, Liu Z, et al. Bioinspired L-Proline oligomers for the cryopreservation of oocytes via controlling ice growth［J］. ACS Appl Mater Interfaces, 2020, 12(16): 18352-18362.
［17］Chi M Q, Chen Z R, Feng Q, et al. L-Proline enhanced whole ovary cryopreservation by inhibiting ice crystal growth and reducing oxidative stress［J］. ACS Biomater Sci Eng, 2025, 11(1): 463-475.
［18］Ruan X Y. Chinese society of gynecological endocrinology affiliated to the international society of gynecological endocrinology guideline for ovarian tissue cryopreservation and transplantation［J］. Gynecol Endocrinol, 2018, 34(12): 1005-1010.
［19］Ruan X, Du J, Korell M, et al. Case report of the first successful cryopreserved ovarian tissue retransplantation in China［J］. Climacteric, 2018, 21(6): 613-616.
［20］Gougeon A. Regulation of ovarian follicular development in primates: facts and hypotheses［J］. Endocr Rev, 1996, 17(2): 121-155.
［21］Ruan X Y, Cui Y M, Du J, et al. Randomized study to prove the quality of human ovarian tissue cryopreservation by xenotransplantation into mice［J］. J Ovarian Res, 2019, 12(1): 46.
［22］Gook D A, Edgar D H, Borg J, et al. Diagnostic assessment of the developmental potential of human cryopreserved ovarian tissue from multiple patients using xenografting［J］. Hum Reprod, 2005, 20(1): 72-78.
［23］Xiao Z, Zhang Y Y, Fan W. Cryopreservation of human ovarian tissue using the silver closed vitrification system［J］. J Assist Reprod Genet, 2017, 34(11): 1435-1444.
［24］Chu Y F, Zhang J L, Wang L Y, et al. The mechanism and protective strategies of follicle injury after ovarian tissue cryopreservation and thawed transplantation: a review［J］. J Ovarian Res, 2025, 18(1): 217.
［25］Gavish Z, Spector I, Peer G, et al. Follicle activation is a significant and immediate cause of follicle loss after ovarian tissue transplantation［J］. J Assist Reprod Genet, 2018, 35(1): 61-69.
［26］Mamsen L S, Olesen H Ø, Pors S E, et al. Effects of Er: YAG laser treatment on re-vascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice［J］. J Assist Reprod Genet, 2021, 38(10): 2745-2756.
［27］Sheshpari S, Shahnazi M, Mobarak H, et al. Ovarian function and reproductive outcome after ovarian tissue transplantation: a systematic review［J］. J Transl Med, 2019, 17(1): 396.

[1]	李凯茵, 阮祥燕, 王泽铖, 张明珍, 柳岸鸣, 王月姣, Alfred O. Mueck. 围绝经期及绝经后女性体成分对骨密度影响的研究[J]. 首都医科大学学报, 2026, 47(4): 680-685.
[2]	张明珍, 阮祥燕, 谷牧青, 杜娟, 程姣姣, Alfred O. Mueck. 医源性早发性卵巢功能不全防治与临床管理指南解读 [J]. 首都医科大学学报, 2026, 47(4): 637-642.
[3]	赵越, 阮祥燕, 王月姣, 谢丹, 金婧, 田玄玄, 张煦, 张娥, 朱建国, 张政. 多囊卵巢综合征患者关键调控分子表达与代谢紊乱的相关性研究[J]. 首都医科大学学报, 2026, 47(4): 643-649.
[4]	李扬璐, 阮祥燕, 程姣姣, 杜娟, 谷牧青, 金凤羽, 李妍秋, 王泽铖, 张明珍, Alfred O. Mueck. 卵巢组织冻存移植后卵泡丢失的机制及N-乙酰半胱氨酸的保护作用研究[J]. 首都医科大学学报, 2026, 47(4): 650-657.
[5]	蒋玲玲, 阮祥燕, 杜娟, 柳岸鸣, 程姣姣, 谷牧青, 李扬璐, 张明珍, 王泽铖, 李妍秋, Alfred O. Mueck. 雌激素替代治疗对裸鼠人卵巢组织异种移植后内分泌功能恢复的影响[J]. 首都医科大学学报, 2026, 47(4): 658-664.
[6]	李妍秋, 阮祥燕, 牧青, 蒋玲玲, 张明珍, Alfred O. Mueck. 屈螺酮炔雌醇片和地屈孕酮治疗对超重多囊卵巢综合征患者体成分与脂代谢的影响[J]. 首都医科大学学报, 2026, 47(4): 665-672.
[7]	郑智, 孙玉君, 李雨筱, 何琳, 侯丽莎, 李琳, 褚春芳. SETX 功能缺失通过影响细胞衰老进而导致早发性卵巢功能不全[J]. 首都医科大学学报, 2026, 47(3): 579-587.
[8]	杨雪, 马宁, 杨玲玲, 黄晓武, 夏恩兰, 李天照. 宫腔粘连与宫腔镜胚物残留术后生育力结局分析：400例单中心回顾性研究[J]. 首都医科大学学报, 2026, 47(2): 360-365.
[9]	宋菁华, 张坤, 郭红燕, 丁慧静. 227例腹壁子宫内膜异位症临床特征分析[J]. 首都医科大学学报, 2026, 47(2): 366-374.
[10]	韩文莉, 邹丽颖, 郑薇, 赵越. 孕早期血红蛋白水平与妊娠期糖尿病发生的关系研究[J]. 首都医科大学学报, 2026, 47(1): 82-86.
[11]	王春雨, 邹丽颖, 赵越. 甲状腺过氧化物酶抗体与游离甲状腺激素对妊娠期糖尿病发病影响的相关性研究[J]. 首都医科大学学报, 2026, 47(1): 87-92.
[12]	兰雪丽, 邹丽颖, 赵越. 妊娠期高血压疾病合并妊娠期糖尿病初产妇不良妊娠结局风险列线图预测模型的构建与验证[J]. 首都医科大学学报, 2026, 47(1): 93-101.
[13]	耿墨钊, 赵越, 邹丽颖. 妊娠期二甲双胍宫内暴露对后代远期健康影响的研究进展[J]. 首都医科大学学报, 2026, 47(1): 108-114.
[14]	戴岩, 桑秀波, 白文佩, 张瑾. 烟酰胺通过降低氧化应激水平缓解不对称二甲基精氨酸诱导的胎鼠宫内生长受限[J]. 首都医科大学学报, 2025, 46(6): 1093-1101.
[15]	王月姣, 阮祥燕, 谷牧青, 魏芸, 管煜伟, 赵越, Alfred O.Mueck. PGRMC1：绝经激素治疗中乳腺癌风险预测的潜在生物学标志物[J]. 首都医科大学学报, 2025, 46(4): 589-593.

天然冷冻保护剂L-脯氨酸与传统冷冻保护剂冻存的人卵巢组织移植有效性研究

Efficacy of the natural cryoprotectant L-proline versus conventional cryoprotectants in human ovarian tissue cryopreservation and transplantation

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价