国际生殖健康/计划生育杂志

国际生殖健康/计划生育 ›› 2018, Vol. 37 ›› Issue (4): 285-287.

• 论著 • 上一篇    下一篇

两种冷冻方法对人卵巢组织冻存效果的分析

张娜，张聪，张辉，贾新转，张轶，唐岚   

  1. 050011  石家庄，河北医科大学第四医院
  • 收稿日期:2018-02-11 修回日期:2018-06-18 出版日期:2018-07-15 发布日期:2018-07-15
  • 基金资助:
    河北省重点研发计划项目（16277766D）

The Cryopreservation Effects of Two Freezing Methods on Human Ovarian Tissue

ZHANG Na, ZHANG Cong, ZHANG Hui, JIA Xin-zhuan, ZHANG Yi, TANG Lan   

  1. The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China
  • Received:2018-02-11 Revised:2018-06-18 Published:2018-07-15 Online:2018-07-15

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摘要： 目的：比较程序化冷冻与玻璃化冷冻人卵巢组织的冻存效果。方法：收集19例人卵巢组织标本，每例分成2份，1份应用程序化方法冷冻，另1份应用玻璃化方法冷冻，复苏后分别移植到去卵巢的雌性裸鼠颈部皮下。38只重症联合免疫缺陷（SCID）裸鼠（移植宿主）随机分为2组：A组19只，移植程序化冷冻复苏的人卵巢组织；B组19只，移植玻璃化冷冻复苏的人卵巢组织。以去除移植宿主双侧卵巢日为D0，移植人卵巢组织日为D21，整个实验过程中，每日通过阴道脱落细胞学检查观察移植宿主动情周期变化，分别于D0、D21、D42、D63取裸鼠静脉血，酶联免疫吸附测定（ELISA）方法检测血清AMH浓度。结果：切除卵巢后移植宿主动情周期消失，D63共有14只裸鼠恢复动情周期，2组间差异无统计学意义（A组8只，B组6只，P＞0.05），但A组恢复时间显著少于B组[（16.38±1.60）d vs.（18.33±1.21）d，P＜0.05]。2组间AMH水平在D0、D21、D42、D63差异均无统计学意义（P＞0.05），但A组、B组D21 AMH水平均显著低于D0，差异有统计学意义（P＜0.05）。恢复动情周期移植宿主的AMH水平在D42和D63逐渐回升但未恢复至D0水平，未恢复动情周期移植宿主的AMH水平在D42和D63均未回升但仍低于D0。结论：程序化冷冻和玻璃化冷冻均能有效保存卵巢组织。在冷冻的卵巢组织经解冻-移植后宿主恢复动情周期方面，程序化冷冻要优先于玻璃化冷冻，但仍需进一步研究。

关键词: 卵巢, 低温保存, 抗苗勒管激素, 程序化冷冻, 玻璃化冷冻, 生育力

Abstract: Objective：To evaluate the cryopreservation effects of the programmed freezing and the vitrification freezing on human ovarian tissue. Methods: Human ovarian tissue specimens of 19 endometrial carcinoma cases were collected. The tissue specimen was divided into two parts: one was cryopreserved with the programmed freezing (the group A), and another with the vitrification freezing （ｔhe group B）． 38 SCID nude mice（transplanted host）were equally divided into two groups，and the thawed human ovarian tissues from the two groups were transplanted to jugular subcutaeous. The day of transplanted host received ovariectomy was defined as D0, while the day of human ovarian tissue transplanted defined as D21. The vaginal cytology examination of transplanted host was performed daily for observation of estrous cycle until D63. The levels of serum AMH on D0, D21, D42 and D63 were measured using ELISA. Results: The estrous cycle of the ovariectomy mice disappeared. A total of 14 transplanted hosts in two groups recovered the estrous cycle up to D63，and there was no significant difference between the two groupｓ （８ in the group A and 6 in the group B, P＞0.0５）． However, the recovering time of the estrous cycle in the group A was significantly shorter than that in the group B [(16.38±1.60) d vs. (18.33±1.21)d, P＜0.05]． There were no significant differences in the AMH levels on D0, D21, D42 and D63 between the two groupｓ （Ｐ＞0.05）． The AMH level on D21 was significantly lower than that on D0 in the group A or B (all P<0.05). The AMH level in those transplanted hosts who had recovered estrous cycle on D42 and D63 was recovered but did not return to the level on D0, while the AMH level in the transplanted hosts who had not recovered their estrous cycle on D42 and D63 did not rebound and was still significantly lower than that on D0. Conclusions: Both the programmed freezing and the vitrification freezing can effectively preserve ovarian tissue.

Key words: Ovary, Cryopreservation, Anti-mullerian hormone, Programmed freezing, Vitrification freezing, Fertility