Fast Freezing Processes in Tissues

组织中的快速冷冻过程

• 批准号: 10200316
• 负责人: Ram Devireddy
• 金额: $ 43.28万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10200316
• 关键词: Area; Authorship; Behavior; Biological; Biological Models; Biomedical Engineering; Calorimetry; Cells; Child; Collagen; Cryopreservation; Cryopreserved Tissue; Cryoprotective Agents; Dermal; Development; Development Plans; Differential Scanning Calorimetry; Discipline; Education; Engineering; Excision; Female; Fibroblasts; Formulation; Freezing; Funding; Gel; Generations; Goals; Grant; Ice; Infrastructure; Instruction; Kinetics; Knowledge; Laboratories; Learning; Learning Module; Liver; Louisiana; Manufacturer Name; Measurement; Measures; Methodology; Methods; Microfabrication; Microscopy; Minority; Modeling; Optics; Participant; Peer Review; Population; Poverty; Process; Protocols documentation; Publications; Research; Resources; Students; System; Techniques; Technology; Testing; Thermodynamics; Time; Tissue Embedding; Tissue Engineering; Tissues; Training; Transplantation; United States National Institutes of Health; Water; Work; base; biological research; biological systems; biophysical properties; cold temperature; design; experimental study; infrastructure development; instrument; laboratory module; neonatal human; novel; outreach; predicting response; programs; recruit; sensor; summer program; symposium; systems research; tool; undergraduate research; undergraduate student

PROJECT SUMMARY The goal of the research is to advance the formulation of cryopreservation protocols for biological systems using fundamental biophysical parameters. At present, crucial gaps exist in our knowledge of the biophysical parameters governing the states of intra- and extra-cellular water during a fast freezing process. This proposal aims to redress this crucial gap by introducing two new and effective techniques: (a) a measurement technique based on microscale thermoelectric sensors, engineered using microfabrication techniques suitable for use with artificial tissues and (b) a measurement technique based on a combination of calorimetry and low temperature microscopy suitable for use with native tissues. Specifically, the project will: (1) design fabricate and test microscale thermoelectric sensors to measure fast freezing processes in isolated cells (Neonatal Human Dermal fibroblasts, NHDFs) and artificial tissues (NHDFs embedded in a collagen gel sheet) with and without Cryoprotective Agents (CPAs) and (2) develop a combination of calorimetric and low temperature microscopy (directional solidification) methods to assess fast freezing processes in NHDFs and native (mammalian liver) tissue sections with and without CPAs. CPA addition and removal in both tissue (artificial and native) systems will be modeled using irreversible thermodynamic models. These integrated methods from disparate disciplines will lead to rational design and development of freezing processes for cryopreservation of donated tissues, an extremely scarce resource, as well as artificially engineered tissues. The education and infrastructure development part of this NIH AREA 15 proposal includes both instructional and research components for LSU undergraduate students.

项目概要 该研究的目标是推进生物系统冷冻保存方案的制定 使用基本的生物物理参数。目前，我们对生物物理学的了解存在重大差距 在快速冷冻过程中控制细胞内和细胞外水状态的参数。这个提议 旨在通过引入两种新的有效技术来弥补这一关键差距：(a) 测量 基于微型热电传感器的技术，采用微加工技术设计 适用于人造组织和 (b) 基于量热法组合的测量技术 以及适用于天然组织的低温显微镜。具体来说，该项目将：（1）设计 制造和测试微型热电传感器以测量隔离细胞中的快速冷冻过程 （新生儿真皮成纤维细胞，NHDF）和人造组织（嵌入胶原凝胶中的 NHDF） (2) 开发量热法和低温法的组合 温度显微镜（定向凝固）方法来评估 NHDF 中的快速冷冻过程和 有或没有 CPA 的天然（哺乳动物肝脏）组织切片。两种组织中 CPA 的添加和去除 （人工和天然）系统将使用不可逆热力学模型进行建模。这些集成的 来自不同学科的方法将导致冷冻工艺的合理设计和开发 冷冻保存极为稀缺的捐赠组织以及人工工程组织。 NIH AREA 15 提案的教育和基础设施发展部分包括教学 以及路易斯安那州立大学本科生的研究部分。