Organizing and Reorganizing Human Testis Development In Vitro

摘要 这是一个多PI、多机构资助项目，旨在开发模仿人类睾丸的仿生系统。 体外发育和生精谱系发育。化疗和放射治疗 对变性人患者进行癌症或性别确认治疗可能会导致不孕。成人患者有 选择在治疗前冷冻卵子或精子，将来可以解冻并用于实现 使用包括试管受精在内的标准辅助生殖技术怀孕。这些选项不适用于 所有成年患者或青春期前不能产生成熟卵子或精子的患者。虽然 接受性别确认治疗的青春期前男孩或变性人女孩不会产生精子，他们会 在他们的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保存未成熟的睾丸组织超过 十年后，人们期待这些组织可以在未来解冻并成熟以产生精子。每个人 患者捐赠一部分组织用于研究开发下一代技术，这将使 它们用自己的组织进行繁殖。我们将使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生命阶段的发育。这将建立基准来评估我们的 三种支持精子发生的新的睾丸仿生系统的体外工程进展。目标1 将使用微流体设备在空气/液体界面复制小鼠睾丸组织器官型培养 在将该方法移植到人类睾丸组织之前，还需要一个静态的PDMS屋顶-转井系统。目标2将 建立小鼠和人的睾丸器官，然后将其诱导融合成细长的 生精小管样结构。AIM 3将使用人类睾丸打印类似生精小管的支架 细胞外基质(HtECM)和猪睾丸ECM(PtECM)水凝胶作为生物墨水，然后种植支架 小鼠和人的睾丸体细胞和生殖细胞。这些睾丸仿生系统将提供 对人类睾丸体细胞和生殖细胞发育的基本见解并建立实验 测试毒理或药物化合物或促进或阻止精子产生的药物的平台。 首要目标是在一个或多个这样的仿生系统中成熟人类睾丸组织。 在体外产生受精能力强的精子。

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.