申请人:Toner Mehmet
公开号:US20050277107A1
公开(公告)日:2005-12-15
The present invention generally relates to devices and methods for the preservation of cells using drying, freezing, and other related techniques. In one set of embodiments, the invention allows for the preservation of cells in a dried state. In another set of embodiments, the invention allows for the preservation of cells within a glass or other non-viscous, non-frozen media. In some embodiments, the invention allows for the preservation of cells at temperatures below the freezing point of water, and in some cases at cryogenic temperatures, without inducing ice formation. The cells, in certain embodiments, may be preserved in the presence of intracellular and/or extracellular carbohydrates (which may be the same or different), for example, trehalose and sucrose. Carbohydrates may be transported intracellularly by any suitable technique, for example, using microinjection, or through non-microinjected methods such as through pore-forming proteins, electroporation, heat shock, etc. In certain instances, the glass transition temperature of the cells may be raised, e.g., by transporting a carbohydrate intracellularly. In some cases, the cells may be dried and/or stored, for example, in a substantially moisture-saturated environment or a desiccating environment. The cells may also be stored in a vacuum or a partial vacuum. The cells may be protected from oxygen, moisture, and/or light during storage. In certain cases, an inhibitor, such as a cell death inhibitor, a protease inhibitor, an apoptosis inhibitor, and/or an oxidative stress inhibitor may be used during preservation of the cells. The cells may be stored for any length of time, then recovered to a viable state, e.g., through rehydration, for further use.
本发明一般涉及利用干燥、冷冻和其他相关技术保存细胞的装置和方法。在一组实施方案中,本发明可以在干燥状态下保存细胞。在另一组实施方案中,本发明可将细胞保存在玻璃或其他非粘性、非冷冻介质中。在某些实施例中,本发明可在低于水的冰点温度下保存细胞,在某些情况下还可在低温下保存细胞,而不会导致冰的形成。在某些实施方案中,细胞可在细胞内和/或细胞外碳水化合物(可相同或不同)的存在下保存,例如,三卤糖和蔗糖。碳水化合物可通过任何合适的技术在细胞内运输,例如使用微量注射,或通过非微量注射方法,如通过孔形成蛋白、电穿孔、热休克等。在某些情况下,可通过在细胞内输送碳水化合物等方法提高细胞的玻璃转化温度。在某些情况下,可将细胞干燥和/或储存,例如,储存在水分基本饱和的环境或干燥环境中。细胞也可以储存在真空或部分真空环境中。在储存过程中,可防止细胞受到氧气、湿气和/或光线的影响。在某些情况下,细胞保存期间可使用抑制剂,如细胞死亡抑制剂、蛋白酶抑制剂、细胞凋亡抑制剂和/或氧化应激抑制剂。细胞可储存任意时间,然后通过复水等方式恢复到可存活状态,以供进一步使用。