BIOENGINEERING PRIMATE FOLLICLES: FROM IMMATURE EGGS TO LIVE BIRTHS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目是题为“肿瘤性生育联盟：为妇女保留生育力”的路线图联盟(U54RR02437/T.Woodruff，PI)的一部分。它的目标是验证在非人类灵长类动物中恢复女性癌症幸存者生育能力的选择，方法是防止卵巢暴露在治疗的配对毒性效应中(通过移除和保存活检)，并返回卵巢组织、配子或胚胎。对猴子的研究：(目的1)生物工程一种支持卵泡三维结构并允许卵泡壁和卵母细胞在体外协调发展的支架；(目的2)评估激素和生长因子在促进卵泡生长和卵母细胞质量方面的作用；(目的3)优化自体卵巢皮质移植的条件，以实现体内协调的卵泡生长和卵母细胞成熟；以及(目的4)评估来自体外成熟卵泡和移植的卵巢皮质卵泡的卵母细胞的受精和胚胎潜能。未成熟的卵泡将从卵巢中分离出来，并在藻酸盐基质中培养。卵巢皮质将被移植到可接近的部位(腹部和前臂)，并将监测促血管生成因子恢复卵巢功能的能力。体外卵泡和皮质移植产生的成熟卵母细胞将通过体外受精、胚胎移植到代孕母亲、怀孕和后代健康等方面进行生殖潜力评估。实验成功地将体外培养30天以上的小卵泡发育到胃窦期，促进类固醇激素(孕激素、雌激素)和旁分泌(抗苗勒氏激素、血管内皮生长因子)功能，并产生体外受精的减数分裂成熟卵。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is part of a Roadmap Consortium (U54RR02437/ T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women". It's goal is to validate options in nonhuman primates for restoring fertility in female cancer survivors by preventing ovarian exposure to the gametotoxic effects of therapy (by removing and preserving biopsies) and returning ovarian tissue, gametes or embryos. Studies in monkeys: (Aim 1) Bioengineer a scaffold that supports the three-dimensional architecture of the follicle and permits coordinated development of the follicle wall and oocyte in vitro; (Aim 2) Evaluate role of hormones and growth factors in promoting follicle growth and oocyte quality; (Aim 3) Optimize conditions for autotransplantation of ovarian cortex for coordinated follicle growth and oocyte maturation in vivo; and (Aim 4) Assess fertilization and embryonic potential of oocytes derived from in vitro matured follicles and transplanted ovarian cortical follicles in vivo. Immature follicles will be isolated from ovaries and cultured in an alginate matrix. Ovarian cortex will be autotransplanted to accessible sites (abdomen and forearm), and the ability of pro-angiogenic factors to restore ovarian function will be monitored. Mature oocytes produced by follicles in vitro and in cortical transplants will be evaluated for reproductive potential by in vitro fertilization, embryo transfer into surrogate mothers, pregnancy and health of offspring. Experiments were successful in growing small follicles for over 30 days in vitro to the antral stage, in promoting steroidogenic (progesterone, estrogen) and paracrine (anti-mullerian hormone, vascular endothelial growth factor) function, and producing meiotically mature eggs that fertilize in vitro.