Towards a preclinical model for overcoming infertility with induced pluripotent stem cells

摘要 在美国，大约12%的女性在怀孕和怀孕方面存在问题， 7%的女性及其伴侣被诊断为不孕不育。一些不孕不育的诊断是可以克服的 然而，通过辅助生殖技术(ART)，这些技术要求每个合作伙伴 功能配子(卵子或精子)才能成功。对于那些不能配子的人来说，艺术就是 不是克服不孕不育诊断的选择。在这笔赠款中，我们正在开发一种生育模型 使用诱导多能干细胞(IPSCs)修复不孕不育患者。这项基础研究建议是 旨在验证产前和新生儿性腺体细胞需要指导男性的假设 以及使用非人灵长类(NHP)iPSCs进行雌性生殖细胞分化。这项建议是 考虑的基础是使用小鼠(M)模型的科学前提，即男性和女性的miPSCs 分化为小鼠原始生殖细胞(PGC)样细胞(MPGCLCs)将经历性别特异性 在与胎儿期性腺体细胞移植时，或直接移植到新生儿的壁龛中时发生分化。这个 这项技术在小鼠中的成功首先建立在对小鼠胚系的基本了解基础上 细胞发育，特别是在胎儿期。在这里，我们提出三个目标，在这些目标中我们将使用 发育生物学、基因组学和移植，以了解性行为的细胞和分子基础- 用NHPs在体内特异性分化PGC。这些知识将被用来区分男性和女性 NHP IPSCs向可通过使用信号逻辑进行性别特异性分化的PGCL分化 产前性腺，或移植后或与产前或新生儿性腺体细胞培养 细胞。在本提案结束时，我们将确定NHP PGCL的程度 能够在产前和新生儿的性腺壁龛中进行性别特异性分化。这项工作将被用来 通知未来的研究，旨在完全从NHP的体外重建男性和女性配子发生。

Summary Around 12% of women in the United States have problems getting pregnant and carrying a baby to term, with 7% of women and their partners being diagnosed as infertile. Some diagnoses of infertility can be overcome through Assisted Reproductive Technologies (ART), however these technologies require each partner to make functional gametes (eggs or sperm) for success. For those individuals incapable of making gametes, ART is not an option for overcoming a diagnosis of infertility. In this grant, we are developing a model for fertility restoration to the infertile patient using induced pluripotent stem cells (iPSCs). This basic research proposal is aimed at testing the hypothesis that prenatal and neonatal gonadal somatic cells are required to instruct male and female germline cell differentiation using non human primate (NHP) iPSCs. The proposal under consideration is based upon the scientific premise with mouse (m) models that male and female miPSCs differentiated into mouse primordial germ cell (PGC)-like cells (mPGCLCs) will undergo sex-specific differentiation when transplanted with prenatal gonadal somatic cells, or directly into neonatal niches. The success of this technology in the mouse was first built upon a fundamental understanding of mouse germline cell development, particularly during prenatal life. Here, we propose three aims in which we will use developmental biology, genomics and transplantation to understand the cell and molecular basis of sex- specific PGC differentiation in vivo using NHPs. This knowledge will be used to differentiate male and female NHP iPSCs towards PGCLCs that can undergo sex-specific differentiation using the signaling logic of the prenatal gonad, or alternatively following transplantation or culture with prenatal or neonatal gonadal somatic cells. At the conclusion of this proposal, we will have determined the extent to which NHP PGCLCs are capable of sex-specific differentiation within prenatal and neonatal gonadal niches. This work will be used to inform future studies aimed at recreating male and female gametogenesis from NHP's entirely in vitro.