Lessons from Nature: Antifreeze Glycolipids for Cryopreservation of Mammalian Cel

大自然的教训：用于哺乳动物细胞冷冻保存的抗冻糖脂

• 批准号: 9035117
• 负责人: LIA H CAMPBELL
• 金额: $ 19.62万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-10 至 2017-07-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035117
• 关键词: Lessons; Nature; Antifreeze; Glycolipids; Cryopreservation

DESCRIPTION (provided by applicant): Storage of cells for many researchers simply involves addition of 10% DMSO or similar cryoprotectants to cells in suspension, putting them in cryovials and slow cooling to subzero temperaturesf and storage in a mechanical or nitrogen cooled freezer or dewar. As long as viable cells are present upon thawing, cell yield may be a secondary consideration due to the high proliferative potential of the cells. However, there are significant cell types that are difficult to preserve with existing cryopreservation techniques including many primary cells, such as cardiac myocytes, and hepatocytes, plus embryonic stem cells and inducible pluripotent stem cells. Others, such as keratinocytes, are easy to cryopreserve in suspension but difficult when attached to a substrate. The study of how organisms survive extreme temperatures has revealed that multiple phenomena may occur in synchrony to promote survival including production of cryoprotectants, antifreeze compounds (peptides and lipids), dehydration and nucleators. Mammalian cell cryopreservation may be possible by mimicking strategies employed by certain insects to survive sub-zero temperatures. These insects combine increases in cryoprotectant content (such as glycerol) with production of antifreeze compounds, at the onset of cold environmental conditions, to survive temperatures as low as -80oC. The presence of these compounds lowers the freezing point of the solution and also changes the shape and formation of ice. In this way, potentially less cryoprotectants, such as DMSO or glycerol, may be used to preserve cells reducing potential cryoprotectant induced cytotoxicity concerns and offering alternative preservation methods for cells and tissues that can be used in clinical applications and cell and tissue-based screening assays for research applications.

描述（由申请人提供）：对于许多研究人员来说，细胞的储存只需要在悬浮液中加入10%的DMSO或类似的冷冻保护剂，将其放入低温瓶中并缓慢冷却至零度以下，然后储存在机械或氮冷却冷冻室或杜瓦瓶中。只要在解冻时存在活细胞，由于细胞的高增殖潜力，细胞产量可能是次要的考虑因素。然而，有一些重要的细胞类型难以用现有的冷冻保存技术保存，包括许多原代细胞，如心肌细胞、肝细胞、胚胎干细胞和诱导多能干细胞。其他细胞，如角质形成细胞，很容易在悬浮液中低温保存，但当附着在底物上时就很难了。对生物如何在极端温度下生存的研究表明，多种现象可能同步发生以促进生存，包括低温保护剂、抗冻化合物（肽和脂质）、脱水和成核剂的产生。通过模仿某些昆虫在零度以下生存的策略，哺乳动物细胞的低温保存可能成为可能。在寒冷的环境条件下，这些昆虫结合了冷冻保护剂含量（如甘油）的增加和防冻化合物的产生，以在低至零下80摄氏度的温度下生存。这些化合物的存在降低了溶液的冰点，也改变了冰的形状和形成。通过这种方式，可能使用较少的冷冻保护剂，如DMSO或甘油，来保存细胞，减少潜在的冷冻保护剂诱导的细胞毒性问题，并提供可用于临床应用的细胞和组织的替代保存方法，以及用于研究应用的基于细胞和组织的筛选分析。