RO1A: Novel Methods for Cryopreservation and Recovery of Human Follicles (2/10)

仅在2006年，预计就有近55 000名育龄前或育龄妇女将被 被诊断为癌症对于接受放疗或化疗治疗癌症的女性，医源性 卵巢损伤是常见的，常常导致不孕。治疗的重大进展， 在过去的几十年中，已经显著提高了被诊断患有癌症的个体的存活率; 5- 1995年至2001年诊断的所有癌症的年相对存活率为65%。据估计 到2010年，每250名成年人中就有1人是癌症幸存者。成效显著 改进的癌症治疗产生了显著的社会需求：即，开发有效的 保持接受这种治疗的个体的生育能力的手段。目前，所有形式的女性生育能力 保存被认为是实验性的。细胞和组织的冷冻保存是一个相互作用的许多 8-12个因素，其中大部分相互作用。通过建立一个明确的理解的基本原则， 细胞或组织的低温生物学特性，我们可以开发数学模型，使我们能够 设计优化方法。在肿瘤生育联盟的赞助下，R 01 A的总体目标是 开发改进的方法来冷冻保存灵长类卵母细胞。未成熟卵泡和卵巢组织，基于 基于合理的基本生物学原理我们建议该项目的具体目标如下： (1)建立并实验性地检验了冷冻保存恒河猴和体外成熟人的优化方法 卵母细胞;（2）：开发和经验性测试冷冻保存恒河猴和人类的优化方法 未成熟卵泡;（3）：开发和经验性测试冷冻保存恒河猴的优化方法， 人类卵巢皮质组织我们提出了一系列的实验来确定细胞的耐受性 对女性生育力至关重要的几个压力与冷冻保存，包括变化， 温度、渗透压和冷冻保护剂。此外，我们还提出了一些实验， 了解卵母细胞，卵泡和卵巢组织的生物物理特性，如渗透性， 细胞对冷冻保护剂的反应以及整个卵巢组织块的细胞内冰形成特性。 结合如上所述的单元对应力的容限使用， 这些信息将允许开发理论上最佳的冷冻保存程序。我们将测试 这些程序使用体外成熟、受精和发育作为实验终点。我们将 最初使用恒河猴卵母细胞、卵泡和卵巢组织进行所有实验。随着新 从这种方法中获得了知识，并开发了用于低温保存的改进方法，我们将 将这些进展应用于人类卵母细胞、卵泡和卵巢组织， 灵长类动物，R 01 C人类卵泡，并结合每一个培训和社会科学 程序（R 01 D、R25、T90/R90、K 01）。

In 2006 alone, nearly 55,000 women either prior to or in their reproductive age range are expected to be diagnosed with cancer. For females undergoing radio- or chemotherapy for cancer treatment, iatrogenic damage to the ovaries is common, often resulting in infertility. Significant advances in treatment during the past few decades have improved the survival significantly among individuals diagnosed with cancer; the 5- year relative survival rate for all cancers diagnosed between 1995 and 2001 is 65%. It has been estimated that; by 2010, 1 in 250 people in the adult population will be cancer survivors. The remarkable success of improved cancer therapy has generated a significant societal need: namely, the development of effective means to preserve the fertility of individuals undergoing such therapies. Currently, all forms of female fertility preservation are considered experimental. Cryopreservation of cells and tissues is an interplay of as many as 8-12 factors, most of which interact. By establishing a clear understanding of the underlying fundamental cryobiological characteristics of the cells or tissues, we can develop mathematical models that enable us to design optimal methods. The overall goal of R01A under the auspices of the Oncofertility Consortium is to develop improved methods to cryopreserve primate oocvtes. immature follicles and ovarian tissue, based upon sound, fundamental crvobiological principles. We propose the following specific aims for this project: (1) develop and empirically test an optimized method for cryopreserving rhesus and human in vitro matured oocytes; (2): develop and empirically test an optimized method for cryopreserving rhesus and human immature follicles; (3): develop and empirically test an optimized method for cryopreserving rhesus and human ovarian cortical tissue. We are proposing a series of experiments to determine the tolerance of cells crucial for female fertility to several stresses associated with Cryopreservation including changes in temperature, osmolality, and cryoprotective agents. In addition, we are proposing experiments designed to understand biophysical characteristics of oocytes, follicles, and ovarian tissue, such as the permeability of the cells to cryoprotectants and the intracellular ice formation characteristics of whole ovarian tissue pieces. Used in conjunction with the tolerances of the cells to the stresses as described above, the resulting information will allow the development of theoretically-optimal Cryopreservation procedures. We will test these procedures using in vitro maturation, fertilization, and development as experimental endpoints. We will initially conduct all of the experiments using rhesus macaque oocytes, follicles and ovarian tissue. As new knowledge is gained from this approach and improved methods for Cryopreservation are developed, we will apply these developments to human oocytes, follicles and ovarian tissue working with the R01B in nonhuman primates, R01C human follicles and in conjuction with each of the training and social science programs (R01D, R25, T90/R90, K01).