促卵泡刺激素干预改善绵羊卵巢组织玻璃化冻存后异种移植的效果研究

背景:卵巢皮质片移植是无血管吻合移植,因此,提高卵巢组织对冻存-解冻和移植后缺血的耐受力是提高冻存后移植物卵泡存活和延长功能寿命的关键环节.目的:观察促卵泡刺激素在玻璃化冻存过程中对绵羊卵巢组织形态和功能的保存效果,为成人卵巢组织的冻存提供技术方法.方法:BALB/c品系雌性裸鼠随机分为3组.均行羊卵巢皮质片裸鼠异位移植:①新鲜移植对照组,取材后即移植.②玻璃化冻存移植组,采用玻璃化冻存解冻后移植,所用液体均未添加促卵泡刺激素.③添加促卵泡刺激素的玻璃化冻存移植组,采用冷冻液、解冻液和培养液均添加促卵泡刺激素的玻璃化冻存后移植.移植后检测受体鼠动情周期恢复率、恢复时间、卵泡数和发育状况,于移植后4周取移植卵巢进行组织学观察、并行血清雌二醇水平分析.结果与结论:含促卵泡刺激素的玻璃化冻存移植组与新鲜移植组相比动情周期恢复率、每高倍视野卵泡计数差异均无显著性意义(P>0.05),卵泡计数高于未添加促卵泡刺激素的玻璃化冻存移植组(P<0.05);动情周期恢复需要的天数接近于新鲜移植组,明显短于未添加促卵泡刺激素的玻璃化冻存移植组(P<0.05).移植后4周,含促卵泡刺激素的玻璃化冻存移植组组织学观察见卵泡发育,未添加促卵泡刺激素的玻璃化冻存移植组仅偶见原始卵泡.含促卵泡刺激素的玻璃化冻存移植组与新鲜移植组相比血清雌二醇水平差异无显著性意义(P>0.05),显著高于未添加促卵泡刺激素的玻璃化冻存移植组(P< 0.05).结果提示,在玻璃化液中加入促卵泡刺激素可提高冻存绵羊卵巢组织移植后卵泡存活率.

Effect of follicle stimulating hormone on cryopreserved sheep ovarian tissue xenotransplantation

BACKGROUND: Transplantation of fragments of ovarian cortex is performed without vascular reanastomosis, therefore, to increase the tolerance of ovarian tissue to the freezing-thawing and ischemic injuries is critical for follicular survival and functional longevity of the graft.OBJECTIVE: To investigate the effect of follicle stimulating hormone (FSH) on the morphological and function of sheep ovary tissue in the process of cryopreservation, so that to provide new freezing method for human ovary tissues. METHODS: Healthy BALB/c strain female nude mice were equally and randomly divided into 3 groups and subjected to heterotopic transplantation of fragments of sheep ovarian cortex. (1) Control group: transplantation following sampling; (2) experimental group: transplantation following freezing-thawing, and the solution was free of FSH; (3) FSH group: the freezing and thawing and culture fluid contained FSH. The estrous cycle recovering rate, estrous cycle recovering time, the number of follicle were observed following transplantation. The histological changes, and the level of E2 in blood serum were observed at 4 weeks after transplantation.RESULTS AND CONCLUSION: There were no significant differences in estrous cycle recovering rate and number of follicle/PHF between FSH and control groups (P> 0.05), but the number of follicle was greater than the experimental group (P< 0.05). Moreover, the time of estrous cycle recovering in FSH group was similar to control group, but shorter than experimental group (P < 0.05). There were more developing follicular in FSH group, but few in experimental group at 4 weeks. No significant difference was detected in E2 level in blood serum between FSH and control group (P > 0.05), but significantly greater than the experimental group (P< 0.05). Results show that FSH addition in vitrification fluid can improve ovarian follicle survival following cryopreserved sheep ovary tissue transplantation.