Kinetics of Intracellular Ice Formation During Cryopreservation of Tissue Engineered Constructs

卡尔松这项提议是对之前工作的扩展，目的是更清楚地定义细胞在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，包括以前所未有的速度(30,000帧/秒)评估IIF的高速视频显微镜，以及利用荧光猝灭测量水传输的新方法(未尝试过)。拟议工作的完成将促进对IIF的启动、生长和传播以及热力学和非热力学变量对这些过程的影响的认识和理解。虽然实验方法使用了双细胞和单层培养，但理论建模有望阐明组织和组织工程化构建物冷冻保存的最佳方法。这项工作包括为研究过程中开发的低温生物学软件实施一个开源项目，其中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。完成这项工作所获得的知识将有益于低温生物学、组织工程和低温外科领域。

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.