Department of Obstetrics and Genecology, University Maternal Hospital, Research Group for Reproductive Medicine and IVF-Laboratory, Cologne University, Cologne, Germany.
Institute of Biology and Immunology of Reproduction, Sofia, Bulgaria.
J Ovarian Res. 2020 Sep 29;13(1):118. doi: 10.1186/s13048-020-00713-9.
The second major cause of death is cancer. In fact, the effectiveness of anticancer treatments and positive long-term prognosis for young women has increased. However, the problem of post-cancer infertility plays a significant role, because chemotherapy can be gonadotoxic and lead to the functional death of ovaries. There is potential key solution to this problem: cryopreservation of ovarian tissue before cancer therapy with re-implantation after convalescence. Data regarding cryopreservation and re-transplantation of ovarian tissue from patients with ovarian insufficiency is limited. The aim of this treatment was the re-transplantation of cryopreserved ovarian tissue after anticancer therapy of patient with ovarian insufficiency (56 IU/l FSH, 8 ng/l β-estradiol, < 1.1 ng/ml anti-Mullerian hormone, 1 primary follicle per 10mm).
After the operation, four tissue fragments (10-16 × 8-13 × 1.0-1.2 mm) were cooled to 5 °C in the freezing medium (culture medium+ 6% ethylene glycol+ 6% dimethyl sulfoxide+ 0.15 M sucrose) for 24 h, frozen and thawed. Freezing was performed in four standard 5 ml cryo-vials with ice formation at - 9 °C, cooling from - 9 to - 34 °C at a rate of - 0.3 °C/min and plunging at - 34 °C into liquid nitrogen. After thawing in a 100 °C (boiling) water bath, the removal of cryoprotectants was performed in 0.5 M sucrose with 20 min. exposure in sucrose and 30 min. stepping rehydration. After thawing of one cryo-vial, part (5 mm) of experimental ovarian tissue after 7 day in vitro culture was histological evaluated and two ovarian fragments (8 × 7 × 1.0 mm and 7 × 6 × 1.0 mm) were re-transplanted. The quantity of follicles after cryopreservation and in vitro culture was not increased (P > 0.1): it was found 1 primordial follicle in 5 mm of tissue. Thirty seven days after the re-transplantation of ovarian tissue, the restoration of the menstrual cycle of Patient W. was noted. Three months after the transplantation, the patient became spontaneously pregnant and delivered a healthy baby girl at term.
Described protocol of conventional cryopreservation of ovarian tissue can be used for treatment of patients with ovarian insufficiency.
癌症是第二大致死原因。事实上，抗癌治疗的效果和年轻女性的积极长期预后有所提高。然而，癌症后不孕的问题仍然存在，因为化疗可能具有性腺毒性，并导致卵巢功能丧失。解决这个问题的关键方法是在癌症治疗前进行卵巢组织冷冻保存，并在康复后进行再植入。关于卵巢功能不全患者的卵巢组织冷冻保存和再移植的数据有限。该治疗的目的是在卵巢功能不全患者（FSH56IU/l、β-雌二醇 8ng/l、抗苗勒管激素<1.1ng/ml、每个 10mm 有 1 个初级卵泡）的抗癌治疗后再移植冷冻保存的卵巢组织。
手术后，将 4 个组织片段（10-16×8-13×1.0-1.2mm）在冷冻介质（培养基+6%乙二醇+6%二甲基亚砜+0.15M 蔗糖）中冷却至 5°C，24 小时，然后冷冻和解冻。在四个标准的 5ml 冷冻管中进行冷冻，在-9°C 形成冰，以 0.3°C/min 的速度从-9°C冷却至-34°C，并在-34°C 下浸入液氮中。解冻后，在 100°C（沸水）水浴中进行 20 分钟的去保护剂处理，然后在蔗糖中暴露 30 分钟进行再水化。解冻一个冷冻管后，对体外培养 7 天后的部分（5mm）实验性卵巢组织进行组织学评估，并再移植两个卵巢组织片段（8×7×1.0mm 和 7×6×1.0mm）。冷冻保存和体外培养后卵泡数量没有增加（P>0.1）：在 5mm 的组织中发现了 1 个原始卵泡。卵巢组织再移植后 37 天，患者 W 的月经周期恢复。移植后 3 个月，患者自然受孕，并足月分娩了一名健康女婴。
所描述的常规卵巢组织冷冻保存方案可用于治疗卵巢功能不全患者。
