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: 10593624
• 负责人: Monica M Laronda
• 金额: $ 20.52万
• 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/10593624
• 关键词: 3-Dimensional; 3D Print; Address; Architecture; Assisted Reproductive Technology; Autologous Transplantation; Biocompatible Materials; Biological; Bioprosthesis device; Cancer Patient; Cattle; Cells; Child; Clinic; Controlled Environment; Cryopreservation; Cryopreserved Tissue; Cues; Development; Disease; Effectiveness; Embryo Transfer; Embryonic Development; Engineering; Environment; Fertility; 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; Mus; Oocytes; Out-Migrations; 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; Seeds; Stromal Cells; Structure; Supporting Cell; Techniques; Technology; Testing; Time; Tissues; Variant; Work; base; 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; 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 >140 例活体 然而，生育力恢复目前仅限于非处方组织移植，部分患者可能会出现这种情况。 由于存在重新引入疾病的风险，无法使用此选项 尚未开发出替代方案。 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）以产生卵子。 技术需要 20 – 30 个卵子才能提供生育孩子的良好机会，但目前的 IVGM 方案。 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有很大不同 在患者之间，最多只有一小部分能够更容易地获得完整的次级卵泡。 保持完整，但在 OTC 期间很少被冷冻保存，因此，必须激活原始卵泡才能生长。 如果使用 OTC 组织来制造卵子，则促进 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 使次级卵泡可靠生长并成熟为优质卵子的 3D 打印生物支架。 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。 结果为进一步探索有希望使用专门设计的生物支架来解决问题奠定了基础 该应用程序测试了动态合成微环境将当前的局限性。 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 3D 打印的卵子。 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 卵泡生长的时间用于模仿人类卵巢的大小、皮质密度、卵泡生长。 并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 这些研究将确定分离的次级卵泡的体外生长和成熟的特定特性。 可以增强卵泡与卵巢组织隔离的合成微环境，以及支持 如果成功的话，这项工作将使研究人员能够研究卵泡的生长和成熟。 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。