Multicomponent solutions for the preservation of cell therapy products

项目摘要 一种新的、受生物启发的策略将被用于改善心肌细胞的无DMSO保存 来源于作为聚集体培养的人类诱导多能干细胞(HIPS)。多细胞系统 对传统保存方法的反应很差。在上一个资金周期中，差异化进化 算法用于使用糖、糖醇和糖的组合来优化细胞的保存 氨基酸。这种方法模仿了植物和其他简单生物的生存策略。 环境压力。表征多组分渗透分子溶液行为的研究 证明了存在于这些溶液中的渗透分子相互作用以改变水的行为 在冷冻过程中，它表现在分子水平上(氢键)，以及 微观结构。渗透分子还与细胞中的关键生物结构相互作用，以稳定它们。低 温度拉曼光谱研究表明，多细胞系统对 过冷(结冰温度和结冰温度之间的温差 在胞外溶液中形成)。当我们从单个细胞过渡到聚集体时，我们再一次看到 向大自然寻求灵感。林蛙使用渗透激素、抑制细胞新陈代谢和 最大限度地减少过冷，因为它适应冬季的冰冻条件。我们正在提出一种类似的方法 以改善心肌细胞聚集体的保存。在申请中描述的研究描述了 IPSC来源的心肌细胞在承诺心脏祖细胞阶段和何时的冷冻反应 形成了完全分化的多细胞心脏结构。过冷度对后融化的影响 完全分化的多细胞心脏结构的恢复也将被确定。的影响 多细胞结构在解冻后恢复时对细胞代谢的抑制也将被量化。vt.在.上 从根本上讲，这些研究将推动我们对更复杂的冻结行为的理解 多细胞系统。在应用层面上，拟议的调查具有保存细胞的潜力 通过使用自然激发的策略从髋关节细胞分化而来。

Project Abstract A novel, biologically inspired strategy will be used to improve DMSO-free preservation of cardiomyocytes derived from human induced pluripotent stem (hiPS) cells cultured as aggregates. Multicellular systems respond poorly to conventional preservation methods. In the previous funding cycle, a differential evolution algorithm was used to optimize the preservation of cells using combinations of sugars, sugar alcohols and amino acids. This approach mimicked the strategies of plants and other simple organisms that survive environmental stresses. Studies characterizing the behavior of multicomponent osmolyte solutions demonstrated that osmolytes present in these solutions interact with each other to modify the behavior of water during freezing which manifests itself at a molecular level (hydrogen bonding) as well as changes in microstructure. The osmolytes also interact with critical biological structures in the cell to stabilize them. Low temperature Raman spectroscopy studies demonstrated that multicellular systems are sensitive to undercooling (the temperature difference between the freezing temperature and the temperature at which ice forms in the extracellular solution). As we transition from single cells to aggregates, once again, we are looking to nature for inspiration. The wood frog uses combinations of osmolytes, suppressed cell metabolism, and minimizes undercooling as it adapts to freezing conditions in the winter. We are proposing a similar approach to improving preservation of cardiomyocyte aggregates. The studies described in the application characterize the freezing response of iPSC-derived cardiomyocytes at the committed cardiac progenitor stage and when formed into fully differentiated, multicellular cardiac constructs. The influence of undercooling on post thaw recovery of fully differentiated multicellular cardiac constructs will also be determined. The influence of suppressing cell metabolism on post thaw recovery of multicellular constructs will also be quantified. On a fundamental level, these studies will advance our understanding of the freezing behavior of more complex multicellular systems. On an applied level, the proposed investigation has the potential to preservation of cells differentiated from hiPS cells through the use of naturally inspired strategies.