Germ Cell Differentiation from Human Embryonic Stem Cells

不孕症困扰着大约10 - 15%的夫妇，其原因与世界上所有的重大事件有关。 生殖途径，从功能性卵子和精子的发育，到受精，胚胎 转录激活、着床、胎盘形成和存活胎儿的生长。尽管频率 不孕症和对受影响者生活质量的影响，很少有人知道潜在的分子和遗传 可使.然而，辅助生殖技术的最新进展，生殖生物学，人类 胚胎干细胞（hESC）生物学和人类遗传学，现在可以让我们克服两个历史性的重大 人类生殖研究的局限性：即人类早期发育的不可及性 到生物探索和人类基因组在发育过程中的遗传难题。在这 我们使用hESC生物学和人类遗传学的强大工具来测试假设：1） 需要DAZ（AZoospermia中的DAZ）基因家族的多个成员来形成和/或 维持新生的人类生殖细胞和2）常见的遗传变异和独特的突变， 在不育男性和女性的染色体中富集，调节两者的早期人类生殖细胞发育。 性别我们的具体目标是：目标1。定义基线分子、时间和遗传特征， hESC和mESC在体外和体内分化为生殖细胞谱系。目标2.沉默人类DAZ， 以及人和小鼠DAZL和BOL基因，并评估生殖细胞发育。目标3。检查功能 DAZ基因家族中常见变异和罕见突变在人群中的意义， 将结果与人类遗传研究和临床发现联系起来。这项研究特别响应了 寻求辅助生殖技术的不孕夫妇的健康问题， 亲生父母事实上，成功完成拟议的研究有望加强我们的 对人类生殖细胞发育的重要途径的基本了解， 为基础科学家研究人类生殖细胞发育，并有助于开发新的 用于临床用途的治疗剂和经验证的诊断基因测试。这项研究将受益匪浅 建立一个中心，提供整个社区的互动和访问有价值的核心 设施本研究使用来自NIH注册的hESC系（UC 01、UC 06、WA 01和WA 09）。

Infertility afflicts approximately 10-15% of couples and has causes linked to all the major events in the reproductive pathways, from development of functional eggs and sperm, to fertilization, embryonic transcriptional activation, implantation, placentation and growth of a viable fetus. In spite of the frequency of infertility and impact on the quality of life of those affected, little is known of underlying molecular and genetic causes. Nonethless, recent advances in assisted reproductive techniques, reproductive biology, human embryonic stem cell (hESC) biology and human genetics, may now allow us to overcome two historicallysignificant limitations in human reproductive studies: namely, the inaccessibility of early human development to biological exploration and the genetic-intractability of the human genome during development. In this proposal, we use the powerful tools of hESC biology and human genetics to test the hypotheses: 1) that multiple members of the DAZ (Deleted in AZoospermia) gene family are required for the formation and/or maintenance of nascent human germ cells and 2) that common genetic variants and unique mutations, enriched in chromosomes of infertile men and women, modulate early human germ cell development in both sexes. Our specific aims are: Aim 1. Define the baseline molecular, temporal and genetic characteristics of differentiation of hESCs and mESCs to the germ cell lineage in vitro and in.vivo. Aim 2. Silence human DAZ, and human and mouse DAZL and BOL genes and assess germ cell development. Aim 3. Examine functional significance of a common variant and a rare mutation in the DAZ gene family in the human population and relate results to human genetic studies and clinical findings. This research is particularly responsive to the health concerns of infertile couples who seek assisted reproductive technologies in hopes of achieving biological parenthood. Indeed, successful completion of the proposed research promises to strengthen our basic understanding of the remarkable pathways by which human germ cells develop, provide useful tools for basic scientists to study human germ cell development, and contribute to the development of novel therapeutics and validated diagnostic genetic tests for clinical use. The research would greatly benefit from the establishment of a center that provides interactions across the community and access to valuable core facilities. This research uses hESC lines from the NIH Registry (UC01, UC06, WA01 and WA09).