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: 7933520
• 负责人: Ralph Lawrence Brinster
• 金额: $ 19.72万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933520
• 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）：精原干细胞（SSC），即雄性生殖系干细胞，通过其自我更新和产生子细胞（分化为精子）的能力，为精子发生提供基础。这些特性使 SSC 成为男性持续生育能力和遗传系保存的关键组成部分。小鼠 SSC 自我更新的信号传导是由生长因子胶质细胞系衍生的神经营养因子 (GDNF) 可能通过 c-Ret 原癌基因受体介导的。使用这种生长因子和特定的培养基，小鼠 SSC 会发生长期（超过 6 个月）体外增殖，这已通过细胞表面抗原标记的维持以及培养的 SSC 在移植到受体时产生供体细胞衍生的精子发生和后代的能力得到证实。因此，一个独特而强大的系统可用于研究小鼠 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群体中GDNF调节的基因，提供可能构成调节SSC自我更新和生存的核心分子机制的基因数据库。其他哺乳动物 SSC 可能使用相同的生长因子和相似的分子机制，因为来自许多物种（例如大鼠、猪、狒狒和人类）的干细胞已被证明可以在小鼠睾丸中增殖。此外，鉴定出的小鼠 SSC 自我更新的重要分子可能有助于理解其他成体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自我更新的分子调控。提出的两个具体目标是：1）研究参与 SSC 自我更新的 GDNF 信号通路，2）确定 GDNF 调节基因在 SSC 自我更新中的生物学意义。鉴定男性生殖系干细胞生长和分裂所需的信号将有助于了解人类男性不育症，并有助于建立在体外培养或生长人类 SSC 的系统。这种培养系统可用于在化疗期间维持男性的干细胞，然后干细胞返回男性的睾丸以在治疗后重建生育能力。