The Russian experience of autotransplantation of vitrified ovarian tissue to a cancer patient

Abstract

To present preliminary results of autotransplantation of vitrified human ovarian tissue to thyroid cancer patient, who underwent radioiodine treatment, performed at MRRC. Cryopreservation of ovarian tissue is increasingly used in the multi-modality treatment of cancer patients. The main purpose of ovarian tissue cryopreservation is to preserve the patient’s fertility before gonadotoxic chemo- and radiotherapy. In 2012, the first orthotopic transplantation of ovarian tissue to the patient previously treated with radioactive iodine was performed at the MRRC in Russia. Monitoring changes in the ovarian reserve allowed us to assess efficacy of the orthotopic transplantation. The results demonstrated a trend to recovery of the patient’s reproductive potential.     

左卡尼汀对玻璃化冷冻小鼠卵母细胞体外受精结局的影响

目的：研究左卡尼汀对玻璃化冷冻小鼠卵母细胞的影响。方法：将卵母细胞分为新鲜组（存活卵数243个）,玻璃化冷冻组（解冻卵数282个）,左卡尼汀高、中、低剂量（2．0、1．0、0．5 g／L）组（解冻卵数分别为285、280和279个）。观察各组卵母细胞中GSH含量、ROS水平以及卵母细胞存活率、受精率、卵裂率、囊胚形成率。结果：5组卵母细胞GSH含量、ROS 水平及卵母细胞存活率、受精率、卵裂率、囊胚形成率差异均有统计学意义（ F ＝881．290、182．760及χ2＝113．361、21．560、16．660、9．667,P均＜0．05）。与新鲜卵母细胞比较,玻璃化冷冻卵母细胞中GSH含量降低,ROS水平升高,卵母细胞存活率、受精率、卵裂率均降低（ P＜0．05）。左卡尼汀呈剂量依赖性地增加玻璃化冷冻卵母细胞中GSH含量,降低ROS水平（P＜0．05）;高剂量左卡尼汀可明显改善玻璃化冷冻卵母细胞的存活率（ P＜0．05）。结论：左卡尼汀可通过抗氧化作用改善玻璃化冷冻卵母细胞的结局。     

Permeability of the plasma membrane to water and cryoprotectants in mammalian oocytes and embryos: Its relevance to vitrification

The permeability of the plasma membrane to water and cryoprotectants is one of the important factors for determining the suitable condition for the vitrification of mammalian oocytes and embryos. Water and cryoprotectants move slowly through oocytes and early embryos, principally by simple diffusion, in the mouse, bovine, pig, and human. In contrast, water, glycerol, and ethylene glycerol move rapidly through morulae and blastocysts, principally by facilitated diffusion via aquaporin 3, in the mouse and bovine; whereas, in the pig, the permeability to water and these cryoprotectants increases not at the morula stage but at the blastocyst stage and further increases at the expanded blastocyst stage. Dimethyl sulfoxide also moves rapidly via channels other than aquaporin 3 in the mouse. In contrast, propylene glycol moves through morulae and blastocysts principally by simple diffusion in the mouse, bovine, and pig, as through oocytes. Therefore, the permeability of mammalian oocytes and embryos at early stages to water and cryoprotectants is low, but that of embryos at later stages to water and some cryoprotectants is markedly high by channel processes, although species specificity exists in some cases.     

Preamble to the proposal for a United Nations second assembly

Maralinga, South Australia was an important site in the United Kingdom nuclear weapons test programme. Seven bombs were exploded and a series of ‘safety’ tests carried out; the latter in particular disseminated appreciable amounts of uranium and plutonium over a wide area. A programme to clean up the test site over several years was instituted, with in-situ vitrification as a principal measure. The final report on the programme has now been published. However, both the programme and the Report are seriously flawed; this article provides criticisms of both.     

Live birth following vitrification of in vitro matured oocytes derived from sibling smaller follicles at follicle selection phase in the context of in vitro fertilization

In ovarian stimulation, a 31-year-old woman with polycystic ovary syndrome was at the risk of developing ovarian hyperstimulation syndrome, follicle aspiration was performed, and eight immature oocytes were collected from follicle fluids. After 28?h in vitro culture, six of them reached MII and were vitrified. The patient failed to conceive in her fresh in vitro fertilization cycle and next two replacement cycles. In the third replacement cycle, a successful pregnancy was obtained by vitrified-thawed oocytes. This case demonstrates that follicular aspiration during follicle selection phase has protective effects against developing ovarian hyperstimulation syndrome, and rescued immature oocytes are viable and could produce promising embryos for live birth.     

Shortened equilibration time can compromise clinical outcomes in human embryo vitrification

Vitrification is an important way to cryopreserve human embryos and the recommended time of embryo exposure to the vitrification solution is 1?min. However, practically speaking, the duration of embryos exposure to equilibration solution can vary from 5 to 15?min. The purpose of this study was to investigate the effect of different equilibration times on the outcomes of frozen-thawed embryo transfer cycles. The data were collected from our medical records from January 2012 to June 2013 and a total of 517 cycles were included. These cycles were divided into four groups according to the equilibration time: (i) 5–6?min; (ii) 7–8?min; (iii) 9–10?min and (iv) 11–12?min. The results show that there were no differences in terms of survival rate and fully intact embryo rate among the four groups. However, lower clinical pregnancy, embryo implantation and live birth rates were observed in the 5–6?min exposure group (54.6%, 31.9% and 48.2%, respectively) compared with the three other groups. The corresponding rates in the 9–10?min group (73.5%, 47.6% and 64.7%) were the highest. This study indicated that different equilibration times influenced the clinical outcomes of human embryo vitrification and vitrification with shortened equilibration time compromised the clinical outcomes. Appropriate prolongation of the equilibrium time would probably improve the clinical outcomes.     

组织玻璃化冻存技术：优势与尚未完全解决的问题

文题释义：玻璃化冷冻保存：是利用这些高浓度的低温保护剂组合成玻璃化冻存液,通过与水分子发生强烈的水合作用,增加溶液黏性,降低冰晶形成速度,从而使细胞在快速降温或复温过程中得以保护。 玻璃化：对于非晶高分子,当高分子通过降温从高弹态转变为玻璃态,或者通过升温从玻璃态转变为高弹态的过程称之为玻璃化转变,发生玻璃化转变的温度叫玻璃化转变温度。对于结晶高分子,玻璃化转变是指其非晶部分所发生的由高弹态向玻璃态(或者玻璃态向高弹态)的转变。因此,玻璃化转变是高分子中普遍存在的现象。但是玻璃化转变现象并不局限于高分子,一些小分子化合物也存在玻璃化转变。 背景：玻璃化冷冻保存是一种应用前途广阔的低温冷冻方法,通过使用高浓度的玻璃化冻存试剂将生物材料进行玻璃态转变,从而实现活性保存。 目的：就玻璃化冻存的生物学原理及玻璃化冻存试剂的分类,卵巢、皮肤与角膜等医学组织标本的玻璃化冻存进行综述。 方法：以“tissue;vitrification;cryopreservation”为英文检索词;“组织;玻璃化;冷冻保存”为中文检索词,检索1994年1月至2019年10月 PubMed 数据库及万方医学网相关文献。按照纳入与排除标准筛选后,对最终纳入的45篇文献进行归纳总结。 结果与结论：玻璃化冻存可以防止细胞内外冰晶形成,避免了冰晶给细胞带来的多种损伤,有效保留了细胞的生物活性与基本功能。玻璃化冻存试剂主要分为渗透性和非渗透性2种,其操作简便、高效,唯一的缺点是高浓度的冻存试剂对细胞具有一定的毒性损伤。为了降低对组织整体损伤风险,可以混合使用多种低毒冻存试剂。目前玻璃化冻存技术已经成功应用于多种细胞,但组织冻存的技术难题尚未完全解决。 ORCID: 0000-0003-0140-9935(张源) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

复苏周期单囊胚移植的影响因素分析

目的研究玻璃化复苏囊胚移植后的妊娠结局与冷冻前囊胚质量、发育速度的相关性,探讨单囊胚移植的可能性。方法回顾分析105例复苏周期单囊胚移植患者的临床资料,观察发育速度、囊胚质量、内细胞团(ICM)/滋养外胚层(TE)评分对妊娠结局的影响。结果第5、6天囊胚间复苏后存活率、移植后妊娠率的差异均无统计学意义(98.0%,54.5%vs 97.0%,52.5%)。优质囊胚移植后妊娠率(44/70,62.9%)显著高于非优质囊胚(12/35,34.3%)。ICM评分为A、B级的囊胚移植后妊娠率差异无统计学意义(70.6%vs 58.0%),但均显著高于C级胚胎(21.1%)。TE评分越低,妊娠率有下降趋势但差异无统计学意义。结论复苏周期单囊胚移植妊娠率与囊胚质量有关,与囊胚发育速度无关。ICM评分而非TE评分与妊娠结果显著相关。复苏周期单个优质囊胚移植后妊娠率高,提示单囊胚移植可行。     

Comparison of two different media for vitrification and rewarming of human zygotes: Prospective randomized study

Vitrification has been successfully used for cryopreservation of human oocytes, zygotes and embryos. There are various media which contain different combinations and concentrations of cryoprotectants for this technique. In this prospective randomized study, two different vitrification and warming media were compared in human zygotes’ cryopreservation.