Kinetics of Intracellular Ice Formation During Cryopreservation of Tissue Engineered Constructs

0541530 Karlsson这一建议是以前工作的延伸，以更清楚地定义细胞物理接触时细胞内和细胞间的冰形成。提出了新的实验技术，包括高速视频显微镜，以前所未有的速度（30，000帧/秒）评估IIF，以及一种新的（未经尝试的）使用荧光猝灭测量水传输的方法。 完成拟议的工作将推进IIF的启动，增长和传播的知识和理解，以及热力学和非热力学变量对这些过程的影响。虽然实验方法使用双细胞和单层培养物，但理论建模有望阐明组织和组织工程构建物冷冻保存的最佳方法。 这项工作包括在研究过程中开发的低温生物学软件的开源项目的实施，包括我们小组以前开发的软件，以及其他研究小组开发的软件。从这项工作的完成中获得的知识将有利于低温生物学，组织工程和冷冻外科领域。

0541530KarlssonThis proposal is an extension of previous work to more clearly define intracellular and intercellular ice formation when cells are physically in contact. New experimental techniques including high speed video microscopy to assess IIF at unprecedented speed (30,000 frames/second), as well as a new (untried) method for measuring water transport using fluorescence quenching are proposed. Completion of the proposed work would advance the knowledge and understanding of IIF initiation, growth, and propagation and the effects of thermodynamic and non-thermodynamic variables on these processes. While the experimental approach uses two-cell and monolayer cultures, theoretical modeling is expected to elucidate optimal approaches for the cryopreservation of tissues and tissue engineered constructs. The work includes an implementation of an open-source project for cryobiology software developed in the course of the research including software previously developed by our group, as well as software developed by other research groups. Knowledge gained from the completion of this work would benefit the field of cryobiology, tissue engineering, and cryosurgery.