Automated High Throughput Cryopreservation Using Microfluidics

使用微流体的自动化高通量冷冻保存

• 批准号: 7802037
• 负责人: Steven Francis Mullen
• 金额: $ 25.1万
• 依托单位: 21ST CENTURY MEDICINE, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2011-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7802037
• 关键词: Archives; Automation; Biological; Biological Preservation; Blood Cells; Cell Culture Techniques; Cells; Chemicals; Chronic Disease; Coupled; Cryopreservation; Cryopreserved Cell; Devices; Diffusion; Effectiveness; Elements; Ethylene Glycols; Excision; Family suidae; Fertility; Goals; Image; Integrated Delivery Systems; Islets of Langerhans Transplantation; Label; Laboratory Animals; Liquid substance; Malignant Neoplasms; Manuals; Medical; Methods; Microfluidics; Microprocessor; Microspheres; Modification; Mus; Oocytes; Ovarian Tissue; Patients; Perfusion; Phase; Phosphate Buffer; Procedures; Process; Property; Pump; Research; Research Personnel; Saline; Sampling; Solutions; Stem cells; Sucrose; Suspension substance; Suspensions; System; Temperature; Therapeutic Uses; Time; Tissues; Toxic effect; Transplantation; Viscosity; aqueous; base; cold temperature; cost; density; design; digital imaging; engineering design; ethylene glycol; improved; instrument; meetings; novel; prototype; public health relevance; research and development; software systems; sperm cell; tool

DESCRIPTION (provided by applicant): The long-term objective of this research is to develop a device that will improve the efficacy of cryopreserving cells in suspension and small pieces of tissue. Having improved means for cell and tissue cryopreservation can increase the effectiveness of bioresource centers containing banks of high quality cells and tissues for research. It can also provide cryopreserved cells for direct patient therapy (e.g. oocytes and ovarian tissue for fertility preservation, stem cells and pancreatic islets for transplantation). The objectives of this Phase I application are to develop a microfluidics cryopreservation system based upon an existing system designed for cell culture and imaging. The current system will be significantly modified in a manner that will allow it to deliver precise concentrations of solutions containing cryoprotectants at temperatures ranging from + 37 to -20 degrees C for cryopreservation of suspended cells. This will facilitate rapid cryoprotectant diffusion into cells before cryopreservation, and dilution out of cells after cryopreservation. It will also allow cells to be exposed to cryoprotectants at reduced temperatures, mitigating chemical toxicity. The aims of this project are to determine means to rapidly mix combinations of solutions and to determine the appropriate flow rates for solutions of varying viscosity. We also plan to design a new microfluidics chip specifically for cryopreservation applications as well as a cooling unit to accompany the existing hardware; a capability that the current device lacks. Mixing will be assessed by analysis of digital images taken of the solution in the final flow path, where metallic microspheres have been suspended into one of the initial solutions. Appropriate flow rates will be determined by assessing the properties of microspheres suspended in the solutions during flow. With such an instrument, high throughput cryopreservation for suspended cells and small tissue can be accomplished, significantly reducing the financial and technical burden associated with cryopreserving laboratory animal germplasm, and also offers a more accurate and reproducible means to archive cells and tissues for research and therapeutic uses. PUBLIC HEALTH RELEVANCE: The proposed research is designed to develop a device that will facilitate making improvements in methods to preserve cells and small pieces of tissue using very low temperatures (cryopreservation). Cryopreserved cells and tissues are important for many reasons, such as direct medical therapy (e.g. transplantation of blood cells), and also for providing consistent biological samples for scientific study by numerous researchers (e.g. when studying cancer or other chronic diseases). Such a device will also significantly increase the efficiency of cryopreserving germplasm from laboratory animals, reducing the overall costs to public agencies for storage and distribution of these valuable research tools.

描述（由申请人提供）：这项研究的长期目的是开发一种设备，该设备将提高冷冻水平细胞在悬浮和小型组织中的功效。改善了细胞和组织冷冻保存的方法可以提高含有高质量细胞和组织库的生物库中心的有效性。它还可以提供冷冻保存的细胞进行直接患者治疗（例如，卵母细胞和卵巢组织可保存生育能力，干细胞和胰岛进行移植）。该阶段应用程序的目标是基于设计用于细胞培养和成像的现有系统开发微流体冷冻保存系统。当前系统将以一种允许其在温度（ + 37至-20摄氏度的温度下）提供精确浓度的溶液的方式进行显着修改，以提供悬浮细胞的冷冻保存。这将促进冷冻保存之前快速的冷冻保护剂扩散到细胞中，并在冷冻保存后从细胞中稀释。它还将使细胞在降低温度下暴露于冷冻保护剂中，从而降低化学毒性。该项目的目的是确定快速混合解决方案组合的方法，并确定适当的粘度溶液流速。我们还计划设计一种专门用于冷冻保存应用的新微流体芯片，以及一个冷却单元，以伴随现有硬件；当前设备缺乏的功能。混合将通过对最终流动路径中溶液拍摄的数字图像的分析进行评估，其中金属微球已被悬浮在初始溶液之一中。适当的流速将通过评估流动过程中溶液中悬浮在溶液中的微球的特性来确定。有了这样的仪器，可以完成用于悬浮细胞和小组织的高吞吐量冷冻保存，从而大大减少与冷冻耐磨实验室动物种质相关的财务和技术负担，还提供了更准确和更可重复的方法来归档细胞和组织，以进行研究和治疗用途。 公共卫生相关性：拟议的研究旨在开发一种设备，以促进使用非常低温（冷冻保存）来保护细胞和小组织的方法。冷冻保存的细胞和组织非常重要，原因有很多，例如直接药物治疗（例如，血液细胞移植），也用于为许多研究人员（例如，研究癌症或其他慢性疾病）提供一致的生物学样品进行科学研究。这样的设备还将显着提高实验动物的冷冻水平种质的效率，从而减少公共机构的总体成本，以存储和分发这些宝贵的研究工具。