Long-term storage does not impact the quality of cryopreserved human ovarian tissue.
Long-term storage does not impact the quality of cryopreserved human ovarian tissue.
复制标题
DOI：
10.1186/s13048-016-0261-8
复制
发表时间：
2016-08-24
影响因子：
4
通讯作者：
Paradisi R
中科院分区：
文献类型：
--
作者：
Fabbri R;Macciocca M;Vicenti R;Pasquinelli G;Caprara G;Valente S;Seracchioli R;Paradisi R
卵巢组织冷冻保存是一项新兴的技术，也适用于非常年轻的癌症患者，对他们来说，在性腺激素治疗之前无法进行卵巢刺激以冷冻卵母细胞。在这种情况下，卵巢组织必须长期冷冻保存，了解它在长期保存后是否保持生育功能是非常重要的。本研究旨在评价长期保存对冷冻保存的人卵巢组织的保存和活力的影响。卵巢组织冷冻保存18年的3例癌症患者的描述性研究对长期保存的组织进行组织学和免疫组织化学分析、透射电子显微镜、原位末端标记法和活/死存活/毒性试验。保存18年的卵巢组织形态良好。卵泡雌激素和孕激素受体阴性，颗粒细胞和/或卵母细胞Ki67阳性，颗粒细胞bcl2阳性。对于间质，雌激素受体和Ki67呈片状/灶状阳性表达，孕激素受体和bcl2呈弥漫性阳性表达。长期保存后，超微结构检查显示滤泡和间质水肿灶的亚细胞完整性。TUNEL法检测未见细胞凋亡。在培养期间，基质细胞的活性保持在97%。对组织不同方面的评估提供了证据，表明储存时间不会影响组织质量，并给癌症女孩带来了希望，她们的组织可以保持很长时间的低温保存。
Ovarian tissue cryopreservation is an emerging technique, also addressed to very young cancer patients, for whom it is not possible to perform an ovarian stimulation for oocytes freezing, before gonadotoxic treatment. In this cases, ovarian tissue must be cryopreserved for a long period of time and it is very important to know if it maintains fertility function after a long period of storage. Here we aimed to assess the effect of long-term storage on preservation and viability of cryopreserved human ovarian tissue. Descriptive study of three cases of cancer patients whose cryopreserved ovarian tissue remained stored for 18 years. Long-term stored tissue was examined by histological and immunohistochemical analysis, transmission electron microscopy, TUNEL assay and LIVE/DEAD viability/citotoxicity test. Ovarian tissue stored for 18 years showed a good morphology. Follicles presented negative staining for estrogen and progesterone receptors, positive staining for ki67 in granulosa cells and/or oocytes and for bcl2 in granulosa cells. Regarding stroma, patch/focal positive expression was found for estrogen receptor and ki67, diffusely positive expression for progesterone receptor and bcl2. After long-term storage, ultrastructural examination showed sub-cellular integrity of follicles and interstitial oedema foci. No apoptosis was observable by TUNEL assay. Stromal cell viability remained >97 % during the culture period. The evaluation of different aspects of the tissue provides evidence that the storage time does not impact on tissue quality and gives hope especially to cancer girls, whose tissues could remain cryopreserved for a very long time.