GENE EXPRESSION IN 3D FOLLICLES

3D 卵泡中的基因表达

• 批准号: 8357851
• 负责人: Mary B Zelinski
• 金额: $ 4.36万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357851
• 关键词: Antral; Biological Preservation; Cancer Survivor; Cell physiology; Cells; Complex; Development; Encapsulated; Female; Fertility; Fertilization; Funding; Gene Expression; Grant; Hormones; Human; Intracytoplasmic Sperm Injections; Knowledge; Life; Macaca; Metaphase; Mus; National Center for Research Resources; Oocytes; Ovulation; Pilot Projects; Primates; Principal Investigator; Process; Research; Research Infrastructure; Resources; Somatic Cell; Source; Staging; Stimulus; System; Technology; Translations; United States National Institutes of Health; Woman; base; cost; embryonic cleavage; folliculogenesis; in vivo; nonhuman primate; offspring; theca cell

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This is a pilot project of the Roadmap Consortium (U54 RR02437, T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women." The process of folliculogenesis in vivo requires the coordinated development and maturation of the follicle wall and the oocyte enclosed within cumulus cells for eventual ovulation and fertilization. The ability to grow mouse follicles in a three-dimensional (3D) culture system yielding mature oocytes capable of fertilization and development to live offspring formed the basis of our current studies in nonhuman primates. We found that macaque follicles can survive, grow, form an antrum and secrete hormones when grown in encapsulated 3D culture. Despite exposure of macaque small antral follicles derived from encapsulated 3D culture to a maturation stimulus, most of the healthy oocytes remained immature. However, two oocytes matured to metaphase II, one fertilized via intracytoplasmic sperm injection and underwent early embryonic cleavage. Encapsulated 3D culture of macaque follicles to the small antral stage allows the functional gene expression of each follicular compartment (somatic cells, oocytes) to be analyzed separately and compared to follicles that develop to an equivalent stage/size in vivo. These studies will assess whether the encapsulated 3D system allows coordinated development of granulosa/theca cells and cumulus-oocyte complexes similar to that in vivo, and if not, will help define cellular functions that require further optimization in the 3D follicle culture system. This knowledge is critical for optimizing the 3D follicle culture technology for translation to human studies which will contribute to fertility preservation for female cancer survivors.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 这是路线图联盟（U 54 RR 02437，T. Woodruff，PI），题为“肿瘤生育联盟：妇女生育力保护。“体内卵泡发生的过程需要卵泡壁和卵丘细胞内的卵母细胞的协调发育和成熟，以最终排卵和受精。在三维（3D）培养系统中生长小鼠卵泡的能力，产生能够受精和发育为活后代的成熟卵母细胞，这是我们目前在非人灵长类动物中研究的基础。我们发现，猕猴卵泡可以存活，生长，形成一个腔，并分泌激素时，在封装的3D培养。 尽管暴露的猕猴小腔卵泡来自封装的3D培养成熟刺激，大多数健康的卵母细胞仍然不成熟。然而，两个卵母细胞成熟到中期II，一个通过卵胞浆内单精子注射受精，并经历早期胚胎卵裂。 将猕猴卵泡包封3D培养至小窦期允许单独分析每个卵泡隔室（体细胞、卵母细胞）的功能基因表达，并与体内发育至等同阶段/大小的卵泡进行比较。这些研究将评估封装的3D系统是否允许颗粒/卵泡膜细胞和卵丘-卵母细胞复合体的协调发育，与体内相似，如果不是，将有助于定义需要在3D卵泡培养系统中进一步优化的细胞功能。这一知识对于优化3D卵泡培养技术以转化为人类研究至关重要，这将有助于女性癌症幸存者的生育力保护。