Centre for Biological Engineering, Dept. of Chemical Engineering, Loughborough University, Loughborough, UK.
Biotechnol Prog. 2011 Nov-Dec;27(6):1511-21. doi: 10.1002/btpr.680.
Human embryonic stem cells (hESCs) and hESC-derived cells are of great interest, not only because of their therapeutic potential but also their prospective uses in in vitro drug and toxicity screening. The ability to preserve these cells is critical, allowing for the generation of quality-controlled stocks of cells, transport of cells between sites, and avoiding the need for expensive and time-consuming continuous culture. Current methodologies, namely conventional slow freezing and vitrification, can successfully preserve hESCs and their differentiated progeny, retaining the key characteristics of the cells. However, there is a significant gap between the number of cells potentially needed to either treat patients or run a high-throughput drug screen and how many cells can be preserved using these techniques.Therefore, this review focuses on the scalability of slow freezing and vitrification, identifying key barriers to success and whether they can be overcome. Given the precedent with other mammalian cells in using slow freezing to successfully preserve large quantities of cells and its compatibility with current and emerging culture methods for hESCs, it is likely to become the method of choice for cryopreserving these cells at scale. However, issues other than scale still exist; therefore, alternatives to cryopreservation should also be explored. Here,the potential to lyophilize hESCs for long-term storage is considered as one such alternative.
人胚胎干细胞（hESCs）和 hESC 衍生细胞备受关注，不仅因为它们具有治疗潜力，还因为它们有望用于体外药物和毒性筛选。保存这些细胞的能力至关重要，可以生成质量可控的细胞库存、在不同地点之间运输细胞，并避免昂贵且耗时的连续培养的需要。目前的方法，即传统的慢速冷冻和玻璃化，能够成功保存 hESCs 及其分化后代，并保留细胞的关键特征。然而，无论是用于治疗患者还是进行高通量药物筛选，所需的细胞数量与可以使用这些技术保存的细胞数量之间存在显著差距。因此，本综述重点关注慢速冷冻和玻璃化的可扩展性，确定成功的关键障碍以及它们是否可以克服。鉴于其他哺乳动物细胞在使用慢速冷冻成功保存大量细胞方面的先例，以及其与 hESC 现有和新兴培养方法的兼容性，慢速冷冻很可能成为大规模冷冻保存这些细胞的首选方法。然而，除了规模之外，仍然存在其他问题；因此，也应该探索冷冻保存以外的替代方法。在这里，考虑将 hESCs 冻干用于长期储存作为一种替代方法。
