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Abstract：
The transplantation of mesenchymal stem cells (MSCs) in patients with premature ovarian failure (POF) could lead to clinical improvement. The transplantation to the ovaries among other transplantation methods have been reported in various animal models, however, there is little evidence regarding the optimal method, including the clinical safety and the efficiency for the treatment of age associated ovarian hypofunction.
To establish the most effective transplantation route of MSCs, explore the resistance to therapy, its safety and role in the natural aging process of the ovaries.
Highly purified MSCs were injected intraperitoneally, directly into the ovaries or tail-intravenously in mice animal model. The ovarian function, quantity and quality of oocytes, cell viability/apoptosis, were evaluated, applying chemiluminescence analysis (CLIA), western blotting, immunofluorescence staining, transmission electron microscope (TEM), TdT mediated dUTP Nick End Labeling (TUNEL) assay and other techniques. The organ tumorigenicity was also evaluated by long-term observation and histopathological examination. The efficiency of MSCs was further verified in non-human primates by the most effective transplantation route.
The 32nd week was ultimately determined as the time point of MSCs transplantation. Our results showed that the intra-ovarian injection was the best transplantation method with a more conspicuous effect. With deeper investigations, we found that the transplanted MSCs showed an effective influence on the follicular number, promoted follicle maturation and inhibited cell apoptosis, which was further verified in non-human primates. In addition, the long-term observation and the histopathological examinations ruled out neoplasms or obvious prosoplasia after MSCs transplantation.
MSCs transplantation by intra-ovarian injection could within a month exert the most conspicuous anti-age-associated ovarian hypofunction effects, which may improve the quantity and quality of oocytes by changing the mitochondrial structure, regulating mitochondrial function and attenuating cell apoptosis to increase the storage of the follicle pool without a remarkable potential of tumorigenicity.
To establish the most effective transplantation route of MSCs, explore the resistance to therapy, its safety and role in the natural aging process of the ovaries.
Highly purified MSCs were injected intraperitoneally, directly into the ovaries or tail-intravenously in mice animal model. The ovarian function, quantity and quality of oocytes, cell viability/apoptosis, were evaluated, applying chemiluminescence analysis (CLIA), western blotting, immunofluorescence staining, transmission electron microscope (TEM), TdT mediated dUTP Nick End Labeling (TUNEL) assay and other techniques. The organ tumorigenicity was also evaluated by long-term observation and histopathological examination. The efficiency of MSCs was further verified in non-human primates by the most effective transplantation route.
The 32nd week was ultimately determined as the time point of MSCs transplantation. Our results showed that the intra-ovarian injection was the best transplantation method with a more conspicuous effect. With deeper investigations, we found that the transplanted MSCs showed an effective influence on the follicular number, promoted follicle maturation and inhibited cell apoptosis, which was further verified in non-human primates. In addition, the long-term observation and the histopathological examinations ruled out neoplasms or obvious prosoplasia after MSCs transplantation.
MSCs transplantation by intra-ovarian injection could within a month exert the most conspicuous anti-age-associated ovarian hypofunction effects, which may improve the quantity and quality of oocytes by changing the mitochondrial structure, regulating mitochondrial function and attenuating cell apoptosis to increase the storage of the follicle pool without a remarkable potential of tumorigenicity.
摘要：
间充质干细胞(MSCs)移植治疗卵巢早衰(POF)患者可改善临床疗效。在各种动物模型中已经报道了除其他移植方法外的卵巢移植。然而,关于最佳方法的证据很少，包括治疗年龄相关性卵巢功能减退的临床安全性和有效性。
建立最有效的骨髓间充质干细胞移植途径，探索对治疗的抵抗力，它在卵巢自然衰老过程中的安全性和作用。
腹膜内注射高度纯化的MSCs，在小鼠动物模型中直接进入卵巢或尾静脉内。卵巢功能,卵母细胞的数量和质量，细胞活力/凋亡，进行了评估，应用化学发光分析(CLIA)，西方印迹,免疫荧光染色，透射电子显微镜（TEM），TdT介导的dUTP尼克末端标记(TUNEL)测定和其他技术。还通过长期观察和组织病理学检查来评估器官致瘤性。通过最有效的移植途径在非人灵长类动物中进一步验证了MSC的效率。
最终确定第32周作为MSCs移植的时间点。我们的结果表明，卵巢内注射是最佳的移植方法，效果更明显。随着更深入的调查，我们发现移植的MSCs对卵泡数量有有效的影响，促进卵泡成熟，抑制细胞凋亡，这在非人灵长类动物中得到了进一步验证。此外,长期观察和组织病理学检查排除了MSCs移植后的肿瘤或明显的前列腺增生。
卵巢内注射MSCs移植可在一个月内发挥最显著的抗年龄相关性卵巢功能减退作用，这可能通过改变线粒体结构来提高卵母细胞的数量和质量，调节线粒体功能和减少细胞凋亡，以增加卵泡池的储存，而没有明显的致瘤潜力。
建立最有效的骨髓间充质干细胞移植途径，探索对治疗的抵抗力，它在卵巢自然衰老过程中的安全性和作用。
腹膜内注射高度纯化的MSCs，在小鼠动物模型中直接进入卵巢或尾静脉内。卵巢功能,卵母细胞的数量和质量，细胞活力/凋亡，进行了评估，应用化学发光分析(CLIA)，西方印迹,免疫荧光染色，透射电子显微镜（TEM），TdT介导的dUTP尼克末端标记(TUNEL)测定和其他技术。还通过长期观察和组织病理学检查来评估器官致瘤性。通过最有效的移植途径在非人灵长类动物中进一步验证了MSC的效率。
最终确定第32周作为MSCs移植的时间点。我们的结果表明，卵巢内注射是最佳的移植方法，效果更明显。随着更深入的调查，我们发现移植的MSCs对卵泡数量有有效的影响，促进卵泡成熟，抑制细胞凋亡，这在非人灵长类动物中得到了进一步验证。此外,长期观察和组织病理学检查排除了MSCs移植后的肿瘤或明显的前列腺增生。
卵巢内注射MSCs移植可在一个月内发挥最显著的抗年龄相关性卵巢功能减退作用，这可能通过改变线粒体结构来提高卵母细胞的数量和质量，调节线粒体功能和减少细胞凋亡，以增加卵泡池的储存，而没有明显的致瘤潜力。
