人类卵巢组织冷冻保存及其评价的研究进展

恶性肿瘤及其治疗过程可能导致女性患者的卵巢功能衰竭,而卵巢组织冷冻保存使这些患者的生育储备以及生殖、内分泌功能的保存成为可能,具有重要的现实意义和广阔的应用前景.目前,卵巢组织冷冻保存技术包括慢速冷冻法、快速冷冻法和玻璃化冷冻法.卵巢组织冻融后主要通过组织学光镜观察卵母细胞和颗粒细胞形态;电镜下观察卵母细胞和间质细胞的线粒体、内质网、核膜和核染色质等超微结构改变评价冻融的影响;冻融组织培养后主要通过观察各卵泡,包括卵泡的直径大小、颗粒细胞的形态和层数、以及卵泡腔和透明带的形成和培养液中激素的含量等评价组织生长发育潜能;通过组织移植技术,检测移植宿主内分泌功能或生殖和内分泌功能的恢复是评价冻存复苏成败的重要指标和目标.     

年轻肿瘤患者冻融卵巢皮质自体移植安全性研究进展

直接冷冻卵巢皮质能保存大量原始卵泡,肿瘤缓解后冻融卵巢皮质自体移植,可以恢复年轻女性患者的生殖功能和内分泌功能,是目前解决癌症患者生育能力最有希望的方法.但是,癌症患者冷冻保存的卵巢组织中可能残留恶性肿瘤细胞,移植后可能导致肿瘤复发.该文综述近年来冻融卵巢皮质自体移植后安全性方面的研究进展.     

冷冻卵巢组织应用的研究进展

癌症治疗过程中放、化疗所造成的医源性月经不规律、卵巢早衰及相关不孕等日益影响到年轻但治愈率高的女性癌症患者的生活质量.卵巢组织的冷冻和移植作为一种新的恢复女性生殖和内分泌功能的手段,越来越受到重视.目前卵巢组织的冷冻方法主要有慢速冷冻法、快速冷冻法和玻璃化冷冻;冻融后卵巢组织的应用途径主要为自体移植、异体移植及未成熟人卵母细胞的体外培养.就卵巢组织的各种冷冻方法和应用途径进行综述,对其目前的研究进展和优缺点进行分析,提出存在的问题.     

冷冻卵巢组织应用的研究进展

癌症治疗过程中放、化疗所造成的医源性月经不规律、卵巢早衰及相关不孕等日益影响到年轻但治愈率高的女性癌症患者的生活质量。卵巢组织的冷冻和移植作为一种新的恢复女性生殖和内分泌功能的手段．越来越受到重视。目前卵巢组织的冷冻方法主要有慢速冷冻法、快速冷冻法和玻璃化冷冻；冻融后卵巢组织的应用途径主要为自体移植、异体移植及未成熟人卵母细胞的体外培养。就卵巢组织的各种冷冻方法和应用途径进行综述，对其目前的研究进展和优缺点进行分析，提出存在的问题。     

人卵巢组织冷冻保存及应用研究进展

卵巢组织冷冻保存可保留年轻的恶性肿瘤和卵巢早衰患者的生殖内分泌功能。目前常用的冷冻方案包括程序化慢速冷冻和玻璃化冷冻。程序化慢速冷冻技术较成熟,已在多领域广泛应用;玻璃化冷冻是近十多年新发展起来的冷冻技术,有研究报道,其冷冻效果优于前者。冻融后卵巢组织主要用于移植和体外培养。卵巢组织自体移植已用于临床,并有成功分娩的报道;体外培养技术尚不成熟,仍处于研究阶段。文献综述人卵巢组织冷冻保存及其应用的研究进展。     

人卵巢组织冷冻保存及应用研究进展

卵巢组织冷冻保存可保留年轻的恶性肿瘤和卵巢早衰患者的生殖内分泌功能.目前常用的冷冻方案包括程序化慢速冷冻和玻璃化冷冻.程序化慢速冷冻技术较成熟,已在多领域广泛应用;玻璃化冷冻是近十多年新发展起来的冷冻技术,有研究报道,其冷冻效果优于前者.冻融后卵巢组织主要用于移植和体外培养.卵巢组织自体移植已用于临床,并有成功分娩的报道;体外培养技术尚不成熟,仍处于研究阶段.文献人卵巢组织冷冻保存及其应用的研究进展.     

人类卵巢组织冷冻方案的研究及其应用前景

人类辅助生殖技术已经作为一种常规的不孕症治疗手段,同时结合精子、胚胎等冷冻技术,使之更好的在人类生殖医学中得到应用。近年来,人们开始致力于卵巢组织冷冻的研究,期望为女性生殖提供保险。不仅如此,卵巢组织的冷冻如果能结合卵巢移植还行望恢复女性内分泌功能,延缓衰老。     

人类卵巢组织冷冻方案的研究及其应用前景

人类辅助生殖技术已经作为一种常规的不孕症治疗手段,同时结合精子、胚胎等冷冻技术,使之更好的在人类生殖医学中得到应用.近年来,人们开始致力于卵巢组织冷冻的研究,期望为女性生殖提供保险.不仅如此,卵巢组织的冷冻如果能结合卵巢移植还有望恢复女性内分泌功能,延缓衰老.     

人类卵巢组织冷冻方案的研究及其应用前景

人类辅助生殖技术已经作为一种常规的不孕症治疗手段，同时结合精子、胚胎等冷冻技术，使之更好的在人类生殖医学中得到应用。近年来，人们开始致力于卵巢组织冷冻的研究，期望为女性生殖提供保险。不仅如此，卵巢组织的冷冻如果能结合卵巢移植还有望恢复女性内分泌功能，延缓衰老。     

家兔卵巢组织异体皮下移植的形态与功能研究

目的:研究家兔卵巢组织玻璃化冷冻、异体皮下移植后激素水平和卵泡的生长发育情况。方法:将36只裸鼠随机分为4组,分别为正常对照组、新鲜移植组、冻存移植组、去势组。将新鲜和冻融卵巢组织分别异体异位移植到裸鼠颈部皮下,于移植术后第5天开始观察裸鼠的动情周期,术后35天左右动情期测鼠血清雌激素水平,并对各组子宫进行湿重及组织形态学观察比较。结果:新鲜及冻存卵巢组织移植后均可观察到存活组织块,移植卵巢内可见不同阶段的发育卵泡,并可见间质腺和黄体。两种移植组于动情期雌激素水平与正常对照组差异无统计学意义(P0.05),均明显高于去势组。结论:玻璃化法冷冻可以有效保存卵巢组织的结构和功能。冻存复苏后与新鲜卵巢皮质组织具有相同的生理活性。冻存卵巢皮质异体异位无血管皮下移植后能够存活、生长发育,并维持生殖内分泌功能。     

Procedures for fertility preservation in female cancer patients

Improving survival rates and quality of life following modern combined cancer treatments have resulted a growing number of patients requesting maintenance of reproductive functions. Several methods are currently available to maintain fertility during oncotherapy. Even though most of them are still experimental and their efficacy and safety have not been determined, the future for fertility preservation in women with cancer is promising. In vitro fertilization with embryo cryopreservation offers an established method, but time to delay cancer treatment could be risky regarding the progression of several cancer types. Moreover, exposure to a high oestrogen milieu during ovarian stimulation is undesirable when patients have estrogen-sensitive malignant tumours. Cryopreservation of mature oocytes following in vitro fertilization and intracytoplasmatic sperm injection offers advantages, but it is still limited due to its low success rate. Emerging techniques of ovarian tissue cryopreservation followed by autotransplantation have been clinically explored. Novel technologies of tissue freezing and thawing promise improving results. However, only one live birth following autotransplanted frozen-thawed ovarian tissue has been established. This procedure can be offered in the future for prepubertal girls before cancer treatment to maintain future fertility. Gonadal tissue cryopreservation and transplantation should be considered experimental in humans for the present time until greater evidence regarding efficacy and safety is accrued.     

卵巢皮质玻璃化冷冻保存及移植的损伤因素

卵巢组织冻存和移植在技术上可行,有效性也随着技术发展而提高,可用于女性生育力保存,尤其是青春期前放化疗的患癌女性的生育能力保存。玻璃化冷冻被认为是目前最有潜力的卵巢组织冻存方法之一,但其安全性和有效性尚需进一步验证。冷冻损伤及移植缺血再灌注损伤是卵巢冻存和移植中需要克服的难题,如何尽可能有效地保存卵泡是难点及重点。从冷冻保护剂、卵巢组织大小、冷冻载体、抗氧化剂、促进新血管形成和激素6个方面对目前已有的玻璃化冷冻及移植条件、效果等进行了阐述,有益于女性生育力保存的项目推进。     

肿瘤生殖学及宫颈癌治疗中生理和生育功能保护问题

随着年轻肿瘤患者的发病率和治愈率均不断提高,以及手术和放化疗常带来不同程度的生育功能损害,保留和保存年轻肿瘤患者的生育功能,成为越来越迫切的需求,肿瘤生殖学(oncofertility)应运而生。肿瘤生殖学是在降低和预防肿瘤复发及由此导致患者死亡的风险,延长患者生存期,甚至治愈癌症的同时,减轻治疗干预方法本身导致的不孕风险,为患者提供选择具有更高生活质量的机会。女性肿瘤患者保存和保护生育功能的主要方法包括：卵巢移位术/性腺屏蔽、在化疗中应用促性腺激素释放激素类似物(GnRHa)保护卵巢功能、胚胎冷冻、卵子冷冻、卵巢组织/卵巢冷冻与移植、干细胞的诱导分化、人工卵巢工程及子宫移植等。宫颈癌治疗中生育功能保留方法包括宫颈广泛切除术(RT)、宫颈锥切术等,此外,保留神经的广泛性子宫切除术和阴道延长术等,也可在宫颈癌治疗中进行生理功能保护,在有效控制肿瘤的同时,使患者生存质量提高。     

接受放化疗年轻女性恶性肿瘤患者的生育能力保存

放疗和化疗对恶性肿瘤的治疗至关重要，但会对年轻女性患者的生育功能造成负面影响，引起不孕。在放化疗前，可以为女性提供多种保留生育能力的选择，包括：卵巢移位、胚胎和卵母细胞冷冻保存、卵巢组织冷冻保存、药物保护性治疗等。将卵巢移位至结肠肝脾曲外侧，可以减少卵巢承受的放射量。卵母细胞和胚胎冷冻保存技术已经成熟，但也有其局限性。卵巢组织冷冻保存目前仍处于试验阶段，已有80多例成功妊娠的案例报道。在化疗期间使用促性腺激素释放激素类似物抑制卵巢功能治疗是一种有争议的保留生育能力的方法。肿瘤医生和生殖医生需要进行多学科合作，权衡保留生育能力与癌症相关的复发率和死亡率，制定个体化治疗方案。     

女性恶性肿瘤患者的生育力保护策略

随着抗肿瘤治疗的发展,癌症患者的生存率大大提高,生育力的保护越来越受到医生和患者的重视。目前,对于女性恶性肿瘤患者,有效的生育力保存方法包括卵子冷冻、胚胎冷冻、卵巢移位术、卵巢组织冷冻与移植和子宫固定术。现就女性恶性肿瘤的生育力保护进行论述。     

冷冻卵巢移植研究进展

哺乳动物的卵巢内有数量众多的卵细胞，是雌性生殖细胞的一个资源库。冷冻保存健康的卵巢组织，快速复苏后再原位移植到盆腔或异位移植至皮下及肾被膜等部位，有望恢复母体的生育力和维持正常激素水平。卵巢组织移植的成功与否取决于移植物的大小、局部缺血持续时间、外源性抗氧化剂和血管生成因子等因素。目前，已有少数关于人类卵巢冷冻移植并成功获得活产儿的报道，也有较多动物实验得到证实。现就冷冻卵巢移植的研究进展进行综述。     

Normal developmental competence to the blastocyst stage is preserved in rabbit ovarian tissue following cryopreservation and autografting to the mesometrium

The aim of the present study was to evaluate mesometrial autotransplantation of frozen-thawed ovarian tissue in the adult rabbit and investigate the developmental competence of oocytes retrieved from grafts by in vitro maturation, fertilisation and blastocyst formation. Twenty-five rabbits were divided into control, fresh tissue transplantation and frozen-thawed tissue transplantation groups. Rabbits were stimulated with follicle-stimulating hormone (FSH) and oocytes were retrieved 3 months after transplantation. Oocytes matured in vivo or in vitro were then fertilised by conventional in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI), followed by observation and evaluation of fertilisation and blastocyst formation rates. No significant differences were found in the percentage of oocytes, maturation, fertilisation, cleavage and blastocyst formation among the three groups. Significantly higher fertilisation rates of in vitro-matured (IVM) oocytes were observed with ICSI compared with IVF in each group (81.1% v. 58.5%, 79.2% v. 59.6% and 80.4% v. 56.0% in the control, fresh tissue transplantation and frozen-thawed tissue transplantation groups, respectively). The blastocyst formation rate of IVM oocytes was significantly lower than that of in vivo-matured oocytes in each group (25.5% v. 65.7%, 22.4% v. 61.8% and 28.9% v. 63.0% in the control, fresh tissue transplantation and frozen-thawed tissue transplantation groups, respectively). In conclusion, the mesometrium is a promising site for ovarian autografts in the rabbit. Oocytes retrieved from mesometrial grafts can develop to the blastocyst stage.     

Assessment of Estrogen Receptors and Apoptotic Factors in Cryopreserved Human Ovarian Cortex

The aims of this study were: (i) to examine frozen-thawed ovarian tissues for features of follicular health and atresia by histology; (ii) to assess the expression of estrogen receptors alpha (ERα) and beta (ERβ) by real-time PCR; (iii) to evaluate the Bax/Bcl-2 ratio, as an apoptotic index, in the ovarian tissues before and after cryopreservation.

Ovarian cortical biopsies were obtained from 11 patients. The fragments were subdivided into two groups, fresh (control tissues) and cryopreserved tissues obtained by direct plunging into liquid nitrogen. Both tissue groups were subjected to a histological evaluation of the healthy and atretic follicles, immunohistochemical localization of the ER, and a real-time PCR (qPCR) to evaluate the expression of ER, Bax, Bcl-2 as well as β-actin, as control gene. Damage was observed in 31% of primordial, 45% of primary, and 75% of secondary follicles in the cryopreserved tissue group. The qPCR analysis showed that the level of ERβ was greater in fresh than cryopreserved tissues, whereas the ERα expression and Bax/Bcl-2 ratio were similar in both tissue groups. A significant inverse association was observed between ERα mRNA levels in the fresh tissue group and subjects' ages.

The results show that cryopreservation and thawing of human ovarian tissue does not affect the morphology of primordial or primary follicles and that cryopreservation does not affect apoptosis. However, cryopreservation seems to have an inhibitory effect on the level of ERβ. Additional studies are needed to evaluate the differential effects of freezing follicles at different stages of follicular development and ovarian steroidogenesis.