Ex Vivo Female Reproductive Tract Integration In a 3D Microphysiologic System

描述（由申请方提供）：女性生殖道负责产生内分泌激素，发育成熟、健康的配子（卵母细胞），并提供受精位点和支持胎儿发育的环境。女性生殖道有五个主要器官：卵巢、输卵管、子宫、子宫颈和阴道。每个器官负责生殖功能的独特方面，但整体上支持整体内分泌健康，生育力和胎儿发育。生殖道器官由多个细胞谱系组装而成，以产生单独的卵泡（包围和支持卵母细胞）、输卵管/输卵管、子宫肌层和子宫内膜、子宫颈和阴道。传统上，女性生殖道的研究依赖于分离的原代细胞或永生化细胞系的二维（2D）培养，这些细胞生长在塑料上，独立于相邻细胞、组织结构和功能背景。转移到三维（3D）培养环境使我们能够更好地了解单个器官内细胞的功能和相互作用，并询问器官中管道组织之间的相互作用。 共培养物（例如，卵泡和卵巢表面细胞，或子宫肌层和子宫内膜），以测量对正常生殖激素、病理条件（如高水平雄激素）或暴露于内分泌干扰物的反应。新的生物材料和3D培养系统现在为我们提供了一个令人兴奋的机会，可以创建一个完整的体外生殖道，从而将每个培养的器官组装成一个连接的灌注培养系统。正如2D单层细胞培养物的生物学功能和反应与3D培养的类器官不同一样，我们预测，在综合系列研究中，生殖器官的生物学将更接近体内环境。在目标1和2中，我们建议开发人类生殖组织的体外培养物，其在激素产生和对生理相关生殖激素促卵泡激素（FSH）和雌激素的反应方面在体内起表型作用。我们将使用3DKUBE”培养平台（KIYATEC），该平台不仅允许控制灌注以模拟组织循环、自动采样以进行药代动力学分析、组织成像和原位生物测定，而且还将有助于将单个器官培养物整合到目标3中的功能性体外女性生殖道培养系统中。离体雌性生殖道的成功开发将使我们能够在完整的生殖道背景下询问每个器官的正常激素反应，以及检查器官和系统对造成生殖危害的药剂的反应。关于雌性生殖功能和生育力的毒理学试验目前仅限于动物研究。我们提出的离体女性生殖道整合在3D微生理系统将允许早期评估药物，毒物或疫苗对人类女性生殖系统的影响之前，在临床试验中暴露。 公共卫生相关性：女性生殖道是一套完整的器官，支持女性的整体内分泌健康，生育力和胎儿发育。管道内的每个器官由相互作用的不同细胞组成，这依赖于精确的组织结构，可以在三维（3D）组织培养中更有效地研究。我们的目标是开发和验证生殖道五个主要器官的3D培养系统，使其成为3D微生理系统中的离体女性生殖道整合，可用于测量对正常激素，内分泌干扰物和其他生殖危害的反应以及药物开发管道。

DESCRIPTION (provided by applicant): The female reproductive tract is responsible for producing endocrine hormones, developing mature, healthy gametes (oocytes) and providing the site for fertilization and an environment that supports fetal development. There are five main organs in the female reproductive tract-the ovary, fallopian tubes, uterus, cervix and vagina. Each organ is responsible for unique aspects of reproductive function, but act integrally to support overall endocrine health, fertility, and fetal development. The reproductive tract organs are assembled from multiple cell lineages to create individual follicles (that enclose and support oocytes), oviductal/fallopian tubes, uterine myometrium and endometrium, the cervix and the vagina. Traditionally, research of the female reproductive tract has relied on two-dimensional (2D) cultures of isolated primary cells or immortalized cell lines grown on plastic and independent of adjacent cells, tissue architecture, and functional context. Moving to a three-dimensional (3D) culture environment has allowed us to better understand the function and interaction of cells within individual organs and interrogate interactions between tract tissues in co-cultures (e.g., the follicle and the ovarian surface cells, or the uterine myometrium and endometrium) to measure responses to normal reproductive hormones, pathologic conditions (such as high levels of androgens) or exposure to endocrine disruptors. New biomaterials and 3D culture systems have now presented us with the exciting opportunity to create a complete in vitro reproductive tract whereby each of the cultured organs can be assembled into a linked perfusion culture system. Just as the biological function and responses of 2D monolayer cell cultures differ from those of 3D-cultured organoids, we predict that the biology of the reproductive organs when studied in an integrated series will more closely recapitulate the in vivo environment. In Aims 1 and 2, we propose to develop in vitro cultures of human reproductive tissues that phenocopy in vivo function in terms of hormone production and response to the physiologically relevant reproductive hormones follicle-stimulating hormone (FSH) and estrogen. We will use the 3DKUBE" culture platform (KIYATEC), which not only permits control of perfusion to mimic tissue circulation, automated sampling for pharmacokinetic analyses, tissue imaging and in situ bioassays, but also will facilitate integration of the individal organ cultures into a functional in vitro female reproductive tract culture system in Aim 3. The successful development of an ex vivo female reproductive tract will give us the unique ability to interrogate normal hormonal responses of each organ in the context of the complete reproductive tract, as well as examine responses of the organs and system to agents that pose reproductive hazards. Toxicologic testing on female reproductive function and fertility is currently limited to animal studies. Our proposed Ex Vivo Female Reproductive Tract Integration In a 3D Microphysiologic System would permit earlier assessment of the effects of drugs, toxicants or vaccines on the human female reproductive system prior to exposure in clinical trials. PUBLIC HEALTH RELEVANCE: The female reproductive tract is an integrated set of organs that supports women's overall endocrine health, fertility and fetal development. Each organ within the tract is composed of different cells that interact with each other, which relies on a precise tissue architecture that can be more effectively studied in three-dimensional (3D) tissue cultures. Our goal is to develop and validate 3D culture systems of the five major organs of the reproductive tract into an Ex Vivo Female Reproductive Tract Integration In a 3D Microphysiologic System that can be used to measure responses to normal hormones, endocrine disruptors, and other reproductive hazards and in the drug development pipeline.