Organizing and Reorganizing Human Testis Development In Vitro

摘要 这是一个多PI，多机构资助，以开发模拟人类睾丸的生理模拟系统 发育和生精谱系发育。化疗和放射治疗， 对变性患者进行癌症或性别确认治疗可能会导致不育。成年患者有 在治疗前冷冻保存卵子或精子的选择，这些卵子或精子可以在将来解冻， 通过标准辅助生殖技术怀孕，包括IVF。这些选项不适用于 所有成年患者或不能产生成熟卵子或精子的青春期前患者。虽然 接受性别确认治疗的青春期前男孩或变性女孩不会产生精子， 睾丸中有精原干细胞，具有产生精子的潜力。中的MPI 芝加哥和匹兹堡已经为年轻患者冷冻未成熟的睾丸组织超过10年。 十年来，人们一直期待这些组织在未来能够解冻并成熟以产生精子。每个 患者捐赠一部分组织用于研究开发下一代技术， 它们利用自己的组织进行繁殖。我们将利用这些组织来获得对人类的基本见解 睾丸从新生儿到成年阶段的发育。这将建立基准来评估我们的 进展工程三种新的睾丸生理模拟系统，支持精子发生，离体。要求1 将使用微流体装置在空气/液体界面处复制小鼠睾丸组织器官型培养 和静态PDMS屋顶-transwell系统，然后将该方法转移到人睾丸组织。目标2将 建立小鼠和人类睾丸类器官，然后诱导其融合成细长的 生精小管样结构。AIM 3将使用人类睾丸打印类似曲细精管的支架 细胞外基质（htECM）和猪睾丸ECM（ptECM）水凝胶作为生物墨水，然后用 小鼠和人类睾丸体细胞和生殖细胞。这些睾丸生理模拟系统将提供 人类睾丸体细胞和生殖细胞发育的基本见解，并建立实验 用于测试促进或阻止精子生成的毒理学或药物化合物或药物的平台。 总体目标是在一个或多个这些生理模拟系统中使人类睾丸组织成熟 体外产生有受精能力的精子。

ABSTRACT This is a multi-PI, multi-institutional grant to develop physiomimetic systems that mimic human testis development and spermatogenic lineage development, ex vivo. Chemotherapy and radiation treatments for cancer or gender affirming treatments for transgender patients can cause infertility. Adult patients have the option to cryopreserve eggs or sperm prior to treatment that can be thawed in the future and used to achieve pregnancy with standard assisted reproductive technologies, including IVF. Those options are not available to all adult patients or to prepubertal patients who are not able to produce mature eggs or sperm. Although prepubertal boys or transgender girls who are on gender affirming treatments are not producing sperm, they do have spermatogonial stem cells SSCs in their testes that have the potential to produce sperm. The MPIs in Chicago and Pittsburgh have been cryopreserving immature testicular tissues for young patients for more than a decade with anticipation that those tissues can be thawed in the future and matured to produce sperm. Each patient donates a portion of their tissue to research to develop next generation technologies that will allow them to use their tissues for reproduction. We will use those tissues to gain fundamental insights into human testis development from newborn to adult stages of life. This will establish benchmarks to evaluate our progress engineering three novel testis physiomimetic systems that support spermatogenesis, ex vivo. Aim 1 will replicate mouse testicular tissue organotypic culture at the air/liquid interface using a microfluidics device and a static PDMS roof-transwell system before translating the approach to human testis tissues. Aim 2 will establish mouse and human testicular organoids, which will then be induced to fuse into elongated seminiferous tubule-like structures. Aim 3 will print seminiferous tubule like scaffolds using human testis extracellular matrix (htECM) and pig testis ECM (ptECM) hydrogels as bioink and then seed the scaffolds with mouse and human testicular somatic cells and germ cells. These testicular physiomimetic systems will provide fundamental insights into human testicular somatic cell and germ cell development and establish experimental platforms to test toxicologic or pharmaceutical compounds or drugs that promote or prevent sperm production. The overarching objective is to mature human testicular tissues in one or more of these physiomimetic systems to produce fertilization competent sperm, ex vivo.