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

仅在2006年，预计将有近55000名育龄前或育龄妇女 被诊断出患有癌症。对于接受放射或化疗治疗癌症的女性，医源性 卵巢受损是很常见的，通常会导致不孕。在治疗过程中取得了重大进展 在过去的几十年里，被诊断为癌症的人的存活率显著提高；5- 1995至2001年间诊断的所有癌症的年相对存活率为65%。据估计， 到2010年，成年人口中每250人中就有1人是癌症幸存者。取得的显著成功 改进的癌症治疗产生了一个重大的社会需求：即开发有效的 保护接受这种治疗的人的生育能力的手段。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许多 作为8-12个因素，其中大部分是相互作用的。通过建立对潜在基本原理的清晰理解 细胞或组织的低温生物学特征，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，R01a的总体目标是 开发冷冻保存灵长类卵子的改进方法。未成熟卵泡和卵巢组织，基于 根据健全的，基本的冰冻生物学原理。我们为这一项目提出了以下具体目标： (1)建立和实验优化恒河猴和人体外成熟冷冻保存方法 卵母细胞；(2)：建立和实验一种优化的恒河猴和人类冷冻保存方法 未成熟卵泡；(3)：开发和实验优化的恒河猴冷冻保存方法和 人卵巢皮质组织。我们提出了一系列实验来确定细胞的耐受性 对雌性生殖力至关重要的几个与冷冻保存相关的压力，包括 温度、渗透压和低温保护剂。此外，我们正在提议进行旨在 了解卵母细胞、卵泡和卵巢组织的生物物理特性，例如 细胞对冷冻保护剂的作用及整个卵巢组织块细胞内结冰的特点。 与如上所述的单元对应力的容差一起使用，结果是 信息将使理论上最佳的冷冻保存程序的发展成为可能。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。我们会 最初，使用猕猴的卵母细胞、卵泡和卵巢组织进行所有的实验。作为新的 从这种方法中获得知识，并开发出改进的冷冻保存方法，我们将 将这些进展应用于使用R01B的人卵母细胞、卵泡和卵巢组织 灵长类，R01C人类卵泡，并与每一个训练和社会科学相结合 程序(R01D、R25、T90/R90、K01)。

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).