Releasing, activating, and maturing follicles from cortical tissue utilizing dynamic synthetic microenvironment

利用动态合成微环境释放、激活和成熟皮质组织中的毛囊

• 批准号: 10710212
• 负责人: Monica M Laronda
• 金额: $ 23.02万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-26 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10710212
• 关键词: 3-Dimensional; 3D Print; Address; Architecture; Assisted Reproductive Technology; Autologous Transplantation; Biocompatible Materials; Biological; Bioprosthesis device; Cancer Patient; Cattle; Cell Aggregation; Cells; Child; Clinic; Controlled Environment; Cryopreservation; Cryopreserved Tissue; Cues; Development; Disease; Effectiveness; Embryo Transfer; Embryonic Development; Engineering; Environment; Fertility; Fertilization; Fertilization in Vitro; Foundations; Freezing; Future; Gelatin; Gender; Goals; Graafian Follicles; Growing Follicle; Growth; Human; Hydrogels; In Situ; In Vitro; Individual; Infertility; Infusion procedures; Left; Live Birth; Mechanics; Methods; Modeling; Mus; Oocytes; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovarian tissue cryopreservation; Ovary; Ovulation; Patients; Porosity; Primordial Follicle; Procedures; Process; Production; Property; Protocols documentation; Reporting; Research; Research Personnel; Risk; Secondary to; Stromal Cells; Structure; Supporting Cell; Techniques; Technology; Testing; Time; Tissues; Variant; Work; bioscaffold; cell motility; childhood cancer survivor; density; design; egg; fertility preservation; folliculogenesis; human tissue; improved; in vivo; migration; novel strategies; offspring; oocyte maturation; oocyte quality; ovary transplantation; patient population; restoration; success; tissue culture; tool; transmasculine; uterine receptivity; zygote

PROJECT SUMMARY/ABSTRACT Ovarian tissue cryopreservation (OTC), a fertility preservation option for cancer patients and others at increased risk of developing infertility that is ideally performed prior to the gonadotoxic treatment, has enabled >140 live births. However, fertility restoration is currently limited to transplantation of OTC tissue, and some patients may not be able to use this option due to the risk of reintroducing their disease. An alternative that is not yet developed for the clinic, would be to use the primordial follicles, the oocyte and support cell units that are cryopreserved in OTC, and perform in vitro growth and maturation (IVGM) to produce eggs. Current assisted reproductive technologies require 20 – 30 eggs to offer a good chance of producing a child. However, current IVGM protocols performed in the research lab are not efficient and yield only a few eggs for every 10 or more patients. Importantly, the success of current methods for isolating primordial follicles from ovarian tissue drastically differs between patients and, at best, only a small percentage are obtained intact. Secondary follicles more easily remain intact, but few are cryopreserved during OTC. Therefore, primordial follicles must be activated to grow to secondary follicles if OTC tissue is used to make eggs. The rate limiting steps for advancing IVGM are: (1) efficiently and reliably isolating healthy primordial follicles that are activated to grow, and (2) efficiently and reliably growing and maturing secondary follicles into good quality eggs. 3D-printed bioscaffolds of specific architectures can support the viability and growth of secondary follicles through egg maturation in vitro, as well as primordial follicle growth and maturation through egg maturation, ovulation, and live birth in mice. These promising results lay the foundation to further explore the use of specifically designed bioscaffolds to address current limitations of IVGM. This application tests the hypothesis that a dynamic synthetic microenvironment will provide the necessary mechanical and architectural cues to induce stromal cell migration out of ovarian tissue, release of the embedded follicles, and support follicle growth and oocyte maturation into an egg. 3D-printed bioscaffolds of varying mechanical and remodeling properties using tunable highly porous biomaterials will be utilized to induce follicle migration from cortical tissue and provide a dynamic environment that remodels over time as the follicles grow. Bovine ovaries as used to mimic human ovaries in size, cortical density, follicle growth and maturation for the following aims: (1) to define the bioscaffolds that support stromal cell migration and subsequent release, activation, and growth of primordial follicles; and (2) to define the bioscaffolds that induce growth and maturation of isolated secondary follicles in vitro. These studies will identify specific properties of synthetic microenvironments that can enhance follicle isolation from ovarian tissue, as well as those that support the growth and maturation of follicles into eggs. If successful, this work would enable researchers to study folliculogenesis from primordial follicle activation to an egg in a controlled environment, would uncover a completely novel approach to IVGM and support the development of future technologies for IVGM in humans.

项目概要/摘要 卵巢组织冷冻保存（OTC）是癌症患者和其他需要增加生育能力的人的生育力保存选择 不孕不育的风险最好在性腺毒性治疗之前进行，已使超过 140 例存活 出生。然而，生育力恢复目前仅限于OTC组织移植，部分患者可能 由于存在重新引入疾病的风险，因此无法使用此选项。尚未开发的替代方案 对于临床，将使用冷冻保存的原始卵泡、卵母细胞和支持细胞单位 OTC，并进行体外生长和成熟（IVGM）以生产鸡蛋。目前辅助生殖 技术需要 20 – 30 个鸡蛋才能提供生育孩子的良好机会。然而，目前的 IVGM 协议 在研究实验室进行的操作效率不高，每 10 名或更多患者只能产下几个卵子。 重要的是，目前从卵巢组织中分离原始卵泡的方法的成功率截然不同 在不同的患者之间，充其量也只有一小部分能够完好无损。次级卵泡更容易 保持完整，但很少有在 OTC 期间冷冻保存。因此，原始卵泡必须被激活才能生长 如果使用 OTC 组织来制造卵子，则为次级卵泡。推进 IVGM 的速率限制步骤是：(1) 有效且可靠地分离被激活生长的健康原始卵泡，以及（2）有效且 使次级卵泡可靠地生长和成熟为优质卵子。 3D打印的特定生物支架 结构还可以通过卵子在体外成熟来支持次级卵泡的活力和生长 小鼠的原始卵泡通过卵子成熟、排卵和活产而生长和成熟。这些 有希望的结果为进一步探索使用专门设计的生物支架来解决问题奠定了基础 IVGM 目前的局限性。该应用程序测试了动态合成微环境将 提供必要的机械和结构线索以诱导基质细胞迁移出卵巢组织， 释放嵌入的卵泡，并支持卵泡生长和卵母细胞成熟为卵子。 3D打印 使用可调节高多孔生物材料的不同机械和重塑特性的生物支架将 用于诱导毛囊从皮质组织迁移并提供重塑的动态环境 随着卵泡生长的时间。牛卵巢在大小、皮质密度、卵泡生长方面模仿人类卵巢 和成熟以实现以下目标：（1）定义支持基质细胞迁移和 随后原始卵泡的释放、激活和生长； (2) 定义诱导的生物支架 体外分离的次级卵泡的生长和成熟。这些研究将确定 可以增强卵泡与卵巢组织隔离的合成微环境，以及支持 卵泡的生长和成熟为卵子。如果成功，这项工作将使研究人员能够研究 在受控环境中从原始卵泡激活到卵子的卵泡发生将揭示 全新的 IVGM 方法，支持人类 IVGM 未来技术的开发。