The commercial and clinical development of cellular therapy products will invariably require cryopreservation and frozen storage of cellular starting materials, intermediates and/or final product. Optimising cryopreservation is as important as optimisation of the cell culture process in obtaining maximum yield and a consistent end-product. Suboptimal cryopreservation can lead not only to batch-to-batch variation, lowered cellular functionality and reduced cell yield, but also to the potential selection of subpopulations with genetic or epigenetic characteristics divergent from the original cell line. Regulatory requirements also impact on cryopreservation as these will require a robust and reproducible approach to the freezing, storage and thawing of the product. This requires attention to all aspects of the application of low temperatures: from the choice of freezing container and cryoprotectant, the cooling rate employed and its mode of delivery, the correct handling of the frozen material during storage and transportation, to the eventual thawing of the product by the end-user. Each of these influences all of the others to a greater or lesser extent and none should be ignored. This paper seeks to provide practical insights and alternative solutions to the technical challenges faced during cryopreservation of cells for use in cellular therapies.

中文翻译：

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用于细胞疗法的干细胞冷冻、低温储存和解冻的技术考虑

细胞治疗产品的商业和临床开发总是需要细胞起始材料、中间体和/或最终产品的冷冻保存和冷冻储存。在获得最大产量和一致的最终产品方面，优化冷冻保存与优化细胞培养过程同样重要。次优冷冻保存不仅会导致批次间变异、细胞功能降低和细胞产量降低，还会导致潜在选择具有与原始细胞系不同的遗传或表观遗传特征的亚群。监管要求也会影响冷冻保存，因为这些要求对产品的冷冻、储存和解冻采用稳健且可重复的方法。这就需要注意低温应用的方方面面：从冷冻容器和冷冻保护剂的选择、采用的冷却速度及其交付方式、在储存和运输过程中正确处理冷冻材料，到最终用户对产品的最终解冻。这些因素中的每一个都或多或少地影响所有其他因素，任何一个都不应该被忽视。本文旨在为用于细胞疗法的细胞冷冻保存过程中面临的技术挑战提供实用的见解和替代解决方案。