New Cell Desiccation Strategy Using Non-Isothermal Drying and Biophysical Models

使用非等温干燥和生物物理模型的新细胞干燥策略

基本信息

批准号：
8989096
负责人：
ALI EROGLU
金额：
$ 19.3万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8989096
关键词：
Address Affect Age Algorithms Area Biological Preservation Biological Sciences Biophysics Cancer Patient Cancer Survivor Cell Membrane Permeability Cell Therapy Cells Chemicals Computer Simulation Containment Cryopreservation Cryoprotective Agents Dehydration Delayed Childbearing Dependence Desiccation Development Diagnosis Diffusion Disease Embryo Emotional Engineering Ethical Issues Exposure to Female Fertility Fertility Agents Fertilization in Vitro Freezing Future Germ Cells Glass Goals Health Human Injury Institutes Legal Life Liquid substance Long-Term Survivors Malignant Neoplasms Mammalian Cell Measurement Measures Medical Methods Mission Modeling Mus National Institute of Biomedical Imaging and Bioengineering Nature Nitrogen Oocytes Organism Outcome Ovarian Hyperstimulation Syndrome Polycystic Ovary Syndrome Polymers Premature Ovarian Failure Preparation Procedures Process Production Property Protocols documentation Quality Control Recording of previous events Recovery Regenerative Medicine Rehydrations Research Research Personnel Residual state Risk Safety Sampling Stem cells Sterility Strategic Planning Stress Techniques Technology Temperature Testing Theoretical model Thermodynamics Time Tissue Engineering Transition Temperature Transplantation Transportation Trehalose Water Woman Work base biobank biophysical model cancer statistics cancer therapy cell injury cell type clinical application cost cytotoxicity design extracellular gonad function improved infertility treatment innovation mathematical model meetings multidisciplinary novel novel strategies programs public health research simulation success sugar young woman

项目摘要

DESCRIPTION: Effective preservation technology for living cells is necessary in biomedical applications ranging from in vitro fertilization to tissue engineering and cell therapies, because the short shelf-life of (non-stabilized) cells imposes severe limitations on biobanking, long-distance transportation, and even safety/quality testing. Current preservation strategies are inadequate, especially for sensitive cell types such as oocytes. For example, cryopreservation requires exposure to high concentrations of penetrating chemicals that are used as cryoprotectants (CPAs) but have inherent cytotoxicity, and requires expensive containment cooled by liquid nitrogen (LN2) for storage and transportation. Moreover, slow-cooling approaches to cryopreservation yield unsatisfactory recovery of healthy oocytes, whereas recent oocyte vitrification approaches require unsafe direct contact with LN2, unreliable sample preparation methods, and much higher CPA concentrations. An alternative preservation strategy, anhydrobiosis (via desiccation), has the potential to stabilize cells at more convenient storage temperatures (e.g., room temperature), but current approaches to desiccation are damaging, because cells are exposed to prolonged osmotic stresses due to diffusion barriers that develop during drying; desiccation of mammalian oocytes is not currently possible. The development of desiccation procedures for human oocytes would allow preservation of fertility for cancer patients, and offer infertility treatment options for other disease states including premature ovarian failure, ovarian hyperstimulation syndrome, and polycystic ovary. The long-term goal of the proposed research program is to meet the critical need of effective, low-cost cell preservation technology by developing a novel, mathematically optimized approach to desiccation. The objective of this developmental (R21) application is to demonstrate feasibility of the proposed method in mouse and human oocytes. This goal will be achieved using a simulation- guided approach, and by taking advantage of unique protective properties of intra- and extracellular sugars. The investigators have previously used this strategy to improve cryopreservation outcome. Informed by preliminary studies indicating that oocytes can be dried to as little as 5% residual moisture content without significant loss of functionality, the central hypothesis is that oocytes can safely be brought into a glassy state at temperatures of -20�C (for storage in a conventional freezer) or 4�C (refrigerator storage) by using a non-isothermal desiccation approach in the presence of intra- and extracellular sugars, sugar polymers, and small amounts (d0.5 M) of a penetrating CPA. The proposed non-isothermal desiccation approach is innovative, because it circumvents the diffusion barrier problem that is inherent to isothermal desiccation, thus allowing faster, less damaging dehydration; other innovations include the development of novel desiccation solutions that use only minimal amounts of CPA, and the use of biophysics-based mathematical models and computer simulations to identify the optimal protocols for non-isothermal desiccation. The proposed research is significant, because its success will (i) shift current paradigms of cell preservation, and (ii) pave the way for meetin biopreservation needs in cell therapy and tissue engineering.

描述：在生物医学应用中，有效的生物细胞保存技术是从体外受精到组织工程和细胞疗法的生物医学应用中，因为（非稳定）细胞的短期遗迹不可能严重限制生物群，长距离运输，甚至是安全/质量测试。当前的保存策略不足，尤其是对于诸如卵母细胞的敏感细胞类型。例如，冷冻保存需要接触高浓度的穿透性化学物质，这些化学物质被用作冷冻保护剂（CPA），但具有固有的细胞毒性，并且需要用液氮（LN2）冷却的昂贵围绕以进行存储和运输。此外，缓慢保存的慢冷却方法导致健康卵母细胞的恢复不佳，而最近的卵母细胞玻璃化方法则需要与LN2，不可靠的样品制备方法和更高的CPA浓度直接直接接触。一种替代的制备策略是赤霉菌病（通过欲望），具有在更方便的储存温度（例如室温）下稳定细胞的潜力，但是当前的相信方法是有害的，因为细胞因在干燥过程中出现的扩散屏障而导致的延长渗透障碍物暴露；目前无法使用哺乳动物卵母细胞。人类卵母细胞的确定程序的开发将允许为癌症患者制备生育能力，并为其他疾病状态提供不孕治疗选择，包括早产卵巢衰竭，卵巢过度刺激综合征和多囊卵巢。拟议的研究计划的长期目标是通过开发一种新颖的，数学优化的确定方法来满足有效的低成本细胞制备技术的关键需求。此发育应用程序的目的（R21）是为了证明小鼠和人卵母细胞中提出的方法。通过模拟引导的方法，并利用细胞内和细胞外糖的独特特性来实现此目标。研究人员以前已经使用该策略来改善冷冻保存结果。通过初步研究得知，表明卵母细胞可以干燥至5％的残留水分含量，而不会显着丧失功能，中央假设是，在-20C的温度（用于存储常规的冰箱中）或4C（冰箱存储）可以通过在存在内和细胞外糖和细胞外糖，糖聚合物，少量（D0.5 m）的情况下，可以安全地将卵母细胞安全地进入玻璃状状态。拟议的非等温欲望方法具有创新性，因为它规避了继承的扩散屏障问题。等温测定，从而允许更快，损害较小的脱水；其他创新包括开发新的确定解决方案，这些解决方案仅使用最少的CPA，以及使用基于生物物理学的数学模型和计算机模拟的使用来确定非等温确定的最佳协议。拟议的研究很重要，因为它的成功将（i）移动电池制备的当前范式，以及（ii）为满足细胞疗法和组织工程中的生物保存需求铺平道路。