IVI Foundation, Valencia, Spain; Reproductive Medicine Research Group; Instituto de Investigación Sanitaria La Fe (IISLAFE), Valencia, Spain.
IVI Foundation, Valencia, Spain; Department of Pediatrics, Obstetrics and Gynecology, School of Medicine, University of Valencia, Valencia, Spain.
Am J Obstet Gynecol. 2021 Jul;225(1):65.e1-65.e14. doi: 10.1016/j.ajog.2021.01.023. Epub 2021 Feb 1.
Ovarian senescence is a normal age-associated phenomenon, but increasingly younger women are affected by diminished ovarian reserves or premature ovarian insufficiency. There is an urgent need for developing therapies to improve ovarian function in these patients. In this context, previous studies suggest that stem cell-secreted factors could have regenerative properties in the ovaries.
This study aimed to test the ability of various human plasma sources, enriched in stem cell-secreted factors, and the mechanisms behind their regenerative properties, to repair ovarian damage and to promote follicular development.
In the first phase, the effects of human plasma enriched in bone marrow stem cell soluble factors by granulocyte colony-stimulating factor mobilization, umbilical cord blood plasma, and their activated forms on ovarian niche, follicle development, and breeding performance were assessed in mouse models of chemotherapy-induced ovarian damage (n=7 per group). In addition, the proteomic profile of each plasma was analyzed to find putative proteins and mechanism involved in their regenerative properties in ovarian tissue. In the second phase, the most effective plasma treatment was validated in human ovarian cortex xenografted in immunodeficient mice (n=4 per group).
Infusion of human plasma enriched bone marrow stem cell soluble factors by granulocyte colony-stimulating factor mobilization or of umbilical cord blood plasma-induced varying degrees of microvessel formation and cell proliferation and reduced apoptosis in ovarian tissue to rescue follicular development and fertility in mouse models of ovarian damage. Plasma activation enhanced these effects. Activated granulocyte colony-stimulating factor plasma was the most potent inducing ovarian rescue in both mice and human ovaries, and proteomic analysis indicated that its effects may be mediated by soluble factors related to cell cycle and apoptosis, gene expression, signal transduction, cell communication, response to stress, and DNA repair of double-strand breaks, the most common form of age-induced damage in oocytes.
Our findings suggested that stem cell-secreted factors present in both granulocyte colony-stimulating factor-mobilized and umbilical cord blood plasma could be an effective treatment for increasing the reproductive outcomes in women with impaired ovarian function owing to several causes. The activated granulocyte colony-stimulating factor plasma, which is already enriched in both stem cell-secreted factors and platelet-enclosed growth factors, seems to be the most promising treatment because of its most potent restorative effects on the ovary together with the autologous source.
卵巢衰老虽是一种与年龄相关的正常现象，但越来越多的年轻女性受到卵巢储备减少或卵巢早衰的影响。因此，迫切需要开发治疗方法来改善这些患者的卵巢功能。在此背景下，先前的研究表明，干细胞分泌的因子可能具有卵巢再生功能。
本研究旨在测试富含干细胞分泌因子的各种人血浆源及其再生特性背后的机制，以修复卵巢损伤并促进卵泡发育。
在第一阶段，通过粒细胞集落刺激因子动员富集骨髓干细胞可溶性因子的人血浆、脐带血血浆及其激活形式对化疗诱导的卵巢损伤的小鼠模型的卵巢巢、卵泡发育和繁殖性能进行评估（每组 7 只）。此外，分析每种血浆的蛋白质组学图谱，以寻找参与其在卵巢组织中再生特性的潜在蛋白和机制。在第二阶段，在免疫缺陷小鼠的人卵巢皮质异种移植中验证最有效的血浆治疗（每组 4 只）。
粒细胞集落刺激因子动员富集骨髓干细胞可溶性因子的人血浆或脐带血血浆的输注导致卵巢组织中微血管形成和细胞增殖程度不同，凋亡减少，从而挽救卵巢损伤模型中的卵泡发育和生育能力。血浆激活增强了这些效果。激活的粒细胞集落刺激因子血浆在小鼠和人卵巢中均具有最强的诱导卵巢挽救作用，蛋白质组学分析表明其作用可能是通过与细胞周期和凋亡、基因表达、信号转导、细胞通讯、应激反应和双链断裂修复相关的可溶性因子介导的，双链断裂是卵母细胞中最常见的与年龄相关的损伤形式。
我们的研究结果表明，富含粒细胞集落刺激因子动员和脐带血血浆中的干细胞分泌因子可能是治疗多种原因导致卵巢功能受损的女性提高生殖结局的有效方法。激活的粒细胞集落刺激因子血浆由于同时富含干细胞分泌因子和血小板封闭生长因子，似乎是最有前途的治疗方法，因为它对卵巢具有最有效的恢复作用，并且是自体来源。
