Regulation of mouse spermatogonial stem cell self-renewal

小鼠精原干细胞自我更新的调控

• 批准号: 7874705
• 负责人: Ralph Lawrence Brinster
• 金额: $ 32.47万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7874705
• 关键词: Adult; Biochemical; Biological; Biological Preservation; Cell Culture System; Cell surface; Cells; Databases; Family suidae; Fertility; Foundations; Genes; Genetic; Germ Lines; Growth Factor; Harvest; Human; In Vitro; Knowledge; Maintenance; Male Infertility; Mediating; Molecular; Molecular Analysis; Mus; Papio; Population; Proliferating; Proto-Oncogenes; Rattus; Regulation; Signal Pathway; Signal Transduction; Spermatogenesis; Stem cell transplant; Stem cells; System; Techniques; Testis; Tissues; Transplantation; chemotherapy; daughter cell; glial cell-line derived neurotrophic factor; male; receptor; self-renewal; sperm cell; stem cell biology

DESCRIPTION (provided by applicant): Spermatogonial stem cells (SSCs), male germ line stem cells, provide the foundation for spermatogenesis by their ability to both self-renew and generate daughter cells, which differentiate into spermatozoa. These attributes make SSCs the key component for sustained fertility in males and the preservation of genetic lines. Signaling for mouse SSC self-renewal is mediated by the growth factor glial cell line-derived neurotrophic factor (GDNF) likely through the c-Ret proto-oncogene receptor. Using this growth factor and a defined medium, long-term (more than 6-months) in vitro proliferation of mouse SSCs occurs, which has been confirmed by maintenance of cell surface antigenic markers and ability of cultured SSCs, when transplanted to recipients, to produce donor cell-derived spermatogenesis and progeny. Thus, a unique and powerful system is available to study the molecular mechanisms of mouse SSC self-renewal. We have recently used microarray profiling to identify GDNF-regulated genes in highly enriched populations of SSCs, providing a database of genes that may constitute the core molecular machinery regulating SSC self-renewal and survival. The same growth factor and similar molecular mechanisms are likely used by other mammalian SSCs, because stem cells from many species (e.g. rat, pig, baboon and human) have been shown to proliferate in mouse testes. In addition, important molecules identified for mouse SSC self-renewal may be helpful in understanding mechanisms used by other adult tissue stem cells. The mouse SSC culture system and stem cell transplantation technique will be used to analyze the molecular regulation of mouse SSC self- renewal. The two proposed specific aims are: 1) Investigate GDNF signaling pathways involved in SSC self- renewal, and 2) Determine the biological significance of GDNF-regulated genes in SSC self-renewal. The identification of the signals required for male germ line stem cells to grow and divide will help in understanding male infertility in humans and in establishing a system to culture or grow human SSCs outside the body. Such a culture system could be used to maintain stem cells of a male during chemotherapy, and the stem cells then returned to the male's testes to reestablish fertility after therapy.

描述（由申请人提供）：雄性干细胞（SSC），雄性生殖系干细胞，通过其自我更新和产生子细胞的能力为精子发生提供了基础，这些能力将其分化为精子。这些属性使SSC成为男性持续生育和保存遗传线的关键组成部分。小鼠SSC自我更新的信号传导可能是通过C-RET原始癌基因受体的生长因子神经胶质细胞系衍生的神经营养因子（GDNF）介导的。使用该生长因子和定义的培养基，发生长期（超过6个月）的小鼠SSC的体外增殖，这已通过维持细胞表面抗原标记和培养的SSC的能力来证实，当将SSC移植到接受者到接受者时，产生供体细胞来源的精子发生和后代。因此，可以使用独特而强大的系统来研究小鼠SSC自我更新的分子机制。我们最近使用微阵列分析来鉴定高度富集的SSC种群中GDNF调节的基因，提供了一个基因数据库，该数据库可能构成调节SSC自我恢复和生存的核心分子机械。其他哺乳动物SSC可能使用了相同的生长因子和相似的分子机制，因为已显示许多物种（例如大鼠，猪，狒狒和人）的干细胞在小鼠睾丸中增殖。此外，针对小鼠SSC自我更新的重要分子可能有助于理解其他成年组织干细胞使用的机制。小鼠SSC培养系统和干细胞移植技术将用于分析小鼠SSC自我更新的分子调节。提出的两个特定目的是：1）研究SSC自我更新中涉及的GDNF信号传导途径，2）确定SSC自我更新中GDNF调节基因的生物学意义。鉴定雄性生殖系干细胞生长和分裂所需的信号将有助于理解男性不孕症，并建立一个培养或在体内培养人类SSC的系统。这种培养系统可用于在化学疗法期间维持雄性的干细胞，然后干细胞返回男性睾丸，以在治疗后重建生育能力。