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个项目摘要 2不育已成为经历过的年轻女孩和妇女的基本生活质量问题 3个性腺毒性癌治疗。一种保留生育能力的突出显示方法正在大量资本化在卵巢中发现的4个原发性卵泡群，而无需自动移植可能是恶性组织。然而， 5在体外开发这些卵泡以产生成熟卵母细胞的成功率效率低下且人类有限 6和非毛碱模型哺乳动物。这项工作的长期目标是建立一种文化体系来支持 7研究牛体外原发性卵泡生长作为人卵泡发生的转化模型。这 8中心假设是使用可降解的聚（乙二醇）（PEG）的仿生培养系统 9可以与中胚层样细胞（MLC）共封存的交联链肽和共囊化 10将更好地促进主要的卵泡发育。这项工作背后的理由是动态的三 11尺寸（3D）培养系统，允许卵泡驱动的基质降解并补充细胞 12类似于卵巢基质（包括丘脑细胞）将概括天然的卵巢环境，更好 13支持长期卵泡发生。该建议的核心目的是检查中胚层的能力 14个细胞（MLC）成为theca样细胞，并促进牛原发性卵泡的发展 15将解离的卵巢细胞纳入PEG水凝胶培养系统中。先前的研究表明馈线 16个细胞，例如脂肪衍生的干细胞，有助于体外的小鼠尿下卵泡的发展。然而， 17 MLC反映了类似于中胚层谱系的细胞身份，这是卵巢的发育起源。 18此外，它们表达卵泡反应性基因，已知对于theca细胞必不可少 19区分和招聘。因此，我们假设它们在脓肿培养中的添加将增加 20创建一个微环境，可以更好地模仿天然卵巢，从而增强对牛的支撑 21卵泡发育。这项工作的新颖方面是一种生物工程文化系统的翻译， 22仅在短期小鼠体外卵泡培养中使用，以使已知可以更好地模拟长期建模的新生物 23和人类卵泡发生的复杂过程。此外，我们在这里测试了源自干细胞的包含 24表达已知对卵泡分泌因子响应的基因创建theca-cell对应物，因此 25进一步强调了该项目的新颖性。这项工作的重要性是它将有助于 26从大型哺乳动物模型物种（如牛）种植原代卵泡的方法的进步， 27将更直接地转化为人类。总体而言，该项目提供了有关使用文化系统和 28补充细胞来源，可以帮助从初级阶段对卵泡发生的整体研究，这样 29我们可以1）更好地了解这个多方面的过程，2）最终创建一个广泛的生育选择选项 30年轻的女孩和女性。