Improving in vitro preantral follicle development using novel bioengineered culture systems and pre-theca-like cells as a strategy for assisted reproduction

使用新型生物工程培养系统和卵泡膜前样细胞作为辅助生殖策略改善体外窦前卵泡发育

基本信息

批准号：
10749434
负责人：
Juliana Candelaria
金额：
$ 4.03万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2024-07-31
项目状态：
已结题

项目摘要

1 PROJECT SUMMARY 2 Infertility has become a fundamental quality-of-life issue for young girls and women who have undergone 3 gonadotoxic cancer treatment. A highlighted approach to preserve fertility is capitalizing on the abundant 4 population of primary follicles found in the ovary without auto-transplanting possibly malignant tissue. However, 5 success rates of developing these follicles in vitro to yield mature oocytes is inefficient and limited in humans 6 and nonmurine model mammals. The long-term goal of this work is to establish a culture system to support the 7 study of in vitro primary follicle growth in the bovine as a translational model for human folliculogenesis. The 8 central hypothesis is that a biomimetic culture system using poly(ethylene glycol) (PEG) with degradable 9 crosslinker peptides and co-encapsulation with mesoderm-like cells (MLCs) that can give rise to theca-like cells 10 will better promote the primary follicle development. The rationale behind this work is that a dynamic three- 11 dimensional (3D) culture system that allows follicle-driven matrix degradation and is supplemented with cells 12 similar to the ovarian stroma (including theca cells) will recapitulate the native ovarian environment and better 13 support long-term folliculogenesis. The central aim of this proposal is to examine the ability of mesoderm-like 14 cells (MLCs) to become theca-like cells and promote development of bovine primary follicles comparable to the 15 inclusion of dissociated ovarian cells in a PEG hydrogel culture system. Previous research has shown that feeder 16 cells, such as adipose-derived stem cells, aid in the development of mouse preantral follicles in vitro. However, 17 MLCs reflect a cell identity similar to the mesoderm lineage, which is the developmental origin of ovary. 18 Additionally, they express follicle-responsiveness genes that are known to be essential for theca cell 19 differentiation and recruitment. Therefore, we hypothesize their addition in preantral follicle culture will add to the 20 creation of a microenvironment that better mimics the natural ovary, thus enhancing support of bovine preantral 21 follicle development. The novel aspect of this work is the translation of a bioengineered culture system, that has 22 only been used in short-term mouse in vitro follicle culture, to a new organism known to better model the long 23 and complex process of human folliculogenesis. Moreover, here we test the inclusion of stemness-derived cells 24 that express genes known to be responsive to follicle-secreted factors to create the theca-cell counterpart, thus 25 further emphasizing the novelty of the project. The significance of this work is that it will contribute to the 26 advancement of methods to grow primary follicles from a large mammal model species like the bovine, which 27 will be more directly translated into human. Overall, this project provides insight on using a culture system and 28 supplemental cell source that can help in the overall study of folliculogenesis from the primary stage such that 29 we can 1) better understand this multifaceted process and 2) ultimately create a broad fertility preservation option 30 for both young girls and women.

1 项目概要 2 不孕不育已成为年轻女孩和妇女的一个基本生活质量问题 3.性腺毒性癌症的治疗。保持生育能力的一个重要方法是利用丰富的资源在卵巢中发现4个初级卵泡群，没有自体移植可能的恶性组织。然而， 5 在体外发育这些卵泡以产生成熟卵母细胞的成功率在人类中效率低下且有限 6和非鼠类模型哺乳动物。这项工作的长远目标是建立支撑文化体系 7 作为人类卵泡发生转化模型的牛体外初级卵泡生长研究。这 8 中心假设是使用具有可降解性的聚乙二醇 (PEG) 的仿生培养系统 9 种交联肽并与中胚层样细胞 (MLC) 共封装，可产生卵泡膜样细胞 10会更好的促进初级卵泡发育。这项工作背后的基本原理是，一个动态的三 11 维 (3D) 培养系统，允许毛囊驱动的基质降解并补充细胞 12 类似于卵巢间质（包括卵泡膜细胞），会更好地再现原生卵巢环境 13 支持长期卵泡发生。该提案的中心目的是检查中胚层样细胞的能力 14 细胞 (MLC) 成为卵泡膜样细胞并促进牛初级卵泡的发育，与 15 将分离的卵巢细胞包含在 PEG 水凝胶培养系统中。先前的研究表明，馈线 16 细胞，例如脂肪干细胞，有助于小鼠腔前卵泡的体外发育。然而， 17 MLC 反映了与中胚层谱系相似的细胞特性，中胚层谱系是卵巢的发育起源。 18 此外，它们还表达已知对卵泡膜细胞至关重要的卵泡反应基因 19 差异化和招聘。因此，我们假设将它们添加到腔前卵泡培养中将增加 20 创建更好地模仿自然卵巢的微环境，从而增强对牛腔前的支持 21 卵泡发育。这项工作的新颖之处在于生物工程培养系统的翻译，该系统具有 22 仅用于短期小鼠体外卵泡培养，以一种已知能更好地模拟长期卵泡的新生物体 23 人类卵泡发生的复杂过程。此外，我们在这里测试了干细胞来源的包含情况 24 表达已知对卵泡分泌因子有反应的基因，以产生卵泡膜细胞对应物，因此 25进一步强调了该项目的新颖性。这项工作的意义在于它将有助于 26 从牛等大型哺乳动物模型物种中培育初级卵泡的方法取得了进展， 27将更直接地翻译成人类。总体而言，该项目提供了有关使用文化系统和 28 种补充细胞来源，可帮助对初级阶段的卵泡发生进行全面研究，以便 29 我们可以 1) 更好地理解这个多方面的过程，2) 最终创造一个广泛的生育力保存选择年轻女孩和妇女均为 30。