Isochoric Pressure Based Preservation of Ovarian Tissue

基于等容压的卵巢组织保存

• 批准号: 9202679
• 负责人: Bradley P Weegman
• 金额: $ 22.5万
• 依托单位: SYLVATICA BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-26 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9202679
• 关键词: Affect; Animals; Biocompatible Materials; Biological; Biological Preservation; Biomedical Engineering; Cancer Patient; Cancer Survivor; Cell Survival; Cell Volumes; Cell model; Cells; Characteristics; Child; Childhood; Climacteric; Clinical; Cryopreservation; Crystallization; Cytoprotection; Development; Dimethyl Sulfoxide; Disease; Effectiveness; Endocrine; Endothelial Cells; Environment; Epithelial; Epithelial Cells; Equilibrium; Excision; Female; Fertility; Freezing; Gold; High temperature of physical object; Human; Ice; Infertility; Injury; Lead; Life; Malignant Neoplasms; Methods; Modeling; Molecular Weight; Nature; Organ; Organ Procurements; Organism; Ovarian; Ovarian Granulosa Cell; Ovarian Tissue; Ovary; Patients; Perfusion; Phase; Phospholipids; Premature Ovarian Failure; Primates; Protocols documentation; Public Health; Research; Resistance; Safety; Series; Staging; Survivors; System; Temperature; Testicular Tissue; Testing; Testis; Thermodynamics; Tissue Banking; Tissue Banks; Tissues; Transplantation; Woman; animation; anticancer research; base; biobank; cancer genomics; cancer therapy; cell type; cold temperature; combinatorial; conditioning; efficacy trial; experience; genomic biomarker; glucose analog; human tissue; implantation; improved; novel; novel strategies; oncofertility; pressure; programs; rate of change; reproductive organ; research clinical testing; solute; success; young adult; young woman

This program aims to develop a novel storage method for human ovarian tissue and whole ovaries, which can also be applied to testicular tissue and gonadal preservation as well as a variety of cells, tissues, and whole organs. This multi-pronged approach builds on recent advances in machine perfusion, nature-inspired cytoprotection strategies, and non-toxic cryoprotectant solutions, combining them with our novel method for constant-volume, pressure- assisted cooling. Our method promises to achieve vitrification of living tissues without the aid of toxic concentrations of cryoprotectants, in a system at thermodynamic equilibrium and potentially at temperatures as high as -70C. It ameliorates or entirely circumvents many of the limitations of conventional cryopreservation methods for large tissues, including damaging ice crystallization, deleterious changes in solute concentrations, and volumetric changes. This approach has the potential to enable indefinite banking of ovarian tissues, whole ovaries, and other living materials while dramatically limiting tissue injury currently associated with ex-vivo storage. The technical objective of this Phase 1 proposal is to develop an optimized cocktail and protocol for preservation and indefinite banking of ovarian tissue strips. We will develop our method across three specific aims. In Aim 1, will use well established methods developed in one of our labs for comprehensive thermodynamic characterization of novel, nature-inspired cryoprotectant solutions in an isochoric (constant-volume) system; this will allow us to select solutions that allow for pressure-assisted vitrification of living materials at low to intermediate pressures and much higher temperatures than traditionally needed for ice-free cryopreservation. In Aim 2, we select the most effective cryoprotectant solutions established in Aim 1 to optimize our “high sub-zero vitrification” in a cell-based model for ovarian tissue consisting of human primary ovarian epithelial cells, human primary ovarian granulosa cells, and human endothelial cells; using high-throughput combinatorial testing, we will select protocols which maximize cell viability and improve on current preservation standards. In Aim 3, we will test our optimized protocols for the preservation of human ovarian cortical tissue strips, along with one of the most successful protocols for ovarian tissue freezing and for ovarian tissue vitrification, respectively; we will additionally test the augmentation of these methods with subnormothermic machine perfusion, enabling further reduction of ischemic tissue injury. Success of these novel approaches individually or in combination will likely enable breakthroughs in clinical oncofertility and biopreservation.

该项目旨在开发一种新的人类卵巢组织和整体储存方法 卵巢，也可应用于睾丸组织和性腺保存以及 这种多管齐下的方法建立在最近的基础上。 机器灌注、自然启发的细胞保护策略和无毒方面的进展 冷冻保护剂解决方案，将其与我们用于恒容、压力的新颖方法相结合 我们的方法有望在不借助辅助冷却的情况下实现活体组织的玻璃化。 在处于热力学平衡的系统中，冷冻保护剂的毒性浓度可能 在高达 -70C 的温度下它改善或完全规避了许多限制。 大组织的传统冷冻保存方法，包括破坏性的冰结晶， 这种方法具有溶质浓度和体积变化的有害变化。 具有无限期储存卵巢组织、整个卵巢和其他活体材料的潜力 同时极大地限制了目前与离体储存相关的组织损伤。 该第一阶段提案的技术目标是开发优化的鸡尾酒和方案 我们将开发我们的方法来保存和无限期保存卵巢组织条。 在目标 1 中，将使用我们实验室之一开发的成熟方法来实现三个具体目标。 新型自然冷冻保护剂的综合热力学表征 等容（恒定体积）系统中的解决方案，这将使我们能够选择允许的解决方案； 用于在低至中压和高压力下对生物材料进行压力辅助玻璃化 在目标 2 中，我们选择比传统无冰冷冻保存所需的温度更高的温度。 目标 1 中建立的最有效的冷冻保护剂解决方案旨在优化我们的“高零度以下 由人原代卵巢上皮组成的基于细胞的卵巢组织模型中的“玻璃化” 使用高通量细胞、人原代卵巢颗粒细胞和人内皮细胞； 组合测试，我们将选择最大化细胞活力并改进当前的方案 在目标 3 中，我们将测试我们优化的保存方案。 人类卵巢皮质组织条，以及最成功的卵巢治疗方案之一 分别用于组织冷冻和卵巢组织玻璃化；我们将另外测试 通过亚常温机器灌注增强这些方法，能够进一步 这些新方法单独或联合治疗的成功。 组合可能会在临床肿瘤生育和生物保存方面取得突破。