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.

项目摘要/摘要 卵巢组织冷冻保存（OTC），癌症患者和其他人的生育能力保存选择 在促性腺毒性治疗之前，理想地进行不孕症的风险已启用> 140个现场 出生。但是，目前，生育能力恢复仅限于OTC组织的移植，某些患者可能 由于有可能重新引入其疾病的风险，因此无法使用此选项。尚未开发的替代方案 对于诊所，将是使用原始卵泡，卵母细胞和支持细胞单位的 OTC并进行体外生长和成熟（IVGM）产生卵。当前的辅助生殖 技术需要20-30个鸡蛋，以提供生育孩子的好机会。但是，当前的IVGM协议 在研究实验室中进行的每10名或更多患者只能产生几个卵。 重要的是，当前方法在卵巢组织中隔离原始卵泡的成功率截然不同 在患者之间，充其量只能获得一小部分。次级卵泡更容易 保持完整，但在OTC期间很少有冷冻保存。因此，必须激活原始卵泡以生长至 次要卵泡如果使用OTC组织制成卵。前进IVGM的限制步骤是：（1） 有效且可靠地隔离活化生长的健康原始卵泡，（2）有效和 可靠地生长和成熟的次要卵泡成高质量的鸡蛋。特定的3D打印生物镜 体系结构也可以通过体外卵成熟来支持次级卵泡的生存能力和生长 作为原始的卵泡生长和通过小鼠排卵和活产的成熟而成熟。这些 有希望的结果为进一步探索专门设计的生物托架的使用奠定了基础 IVGM的当前局限性。该应用程序检验了一个动态合成微环境的假设 提供必要的机械和建筑线索，以诱导基质细胞从卵巢组织中迁移， 释放嵌入的卵泡，并支持卵泡生长和卵母细胞成熟到卵中。 3D打印 使用可调的高度多孔生物材料的不同机械和重塑特性的生物施加物将是 用于诱导卵泡从皮质组织迁移，并提供一个动态环境，可以重塑 随着follices的增长时间。用于模仿大小，皮质密度，卵泡生长的牛卵巢 和以下目的的成熟：（1）定义支持基质细胞迁移和 原始卵泡的随后释放，激活和生长； （2）定义影响 在体外分离的次生卵泡的生长和成熟。这些研究将确定 可以增强卵巢组织分离的叶分离的合成微环境，以及支持的叶面微环境 卵泡的生长和成熟到卵中。如果成功，这项工作将使研究人员能够学习 在受控环境中，从原始叶子激活到卵的卵泡发生，会发现 IVGM的完全新颖的方法并支持人类IVGM未来技术的发展。