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首页> 外文期刊>Biopreservation and biobanking >Toward Optimal Cryopreservation and Storage for Achievement of High Cell Recovery and Maintenance of Cell Viability and T Cell Functionality
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Toward Optimal Cryopreservation and Storage for Achievement of High Cell Recovery and Maintenance of Cell Viability and T Cell Functionality

机译：寻求最佳的冷冻保存方式以实现高细胞回收率并维持细胞活力和T细胞功能

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摘要

Cryopreservation of biological materials such as cells, tissues, and organs is a prevailing topic of high importance. It is employed not only in many research fields but also in the clinical area. Cryopreservation is of great importance for reproductive medicine and clinical studies, as well as for the development of vaccines. Peripheral blood mononuclear cells (PBMCs) are commonly used in vaccine research where comparable and reliable results between different research institutions and laboratories are of high importance. Whereas freezing and thawing processes are well studied, controlled, and standardized, storage conditions are often disregarded. To close this gap, we investigated the influence of suboptimal storage conditions during low-temperature storage on PBMC viability, recovery, and T cell functionality. For this purpose, PBMCs were isolated and exposed with help of a robotic system in a low-temperature environment from 0 up to 350 temperature fluctuation cycles in steps of 50 cycles to simulate storage conditions in large biorepositories with sample storage, removal, and sorting functions. After the simulation, the viability, recovery, and T cell functionality were analyzed to determine the number of temperature rises, which ultimately lead to significant cell damage. All studied parameters decreased with increasing number of temperature cycles. Sometimes after as little as only 50 temperature cycles, a significant effect was observed. These results are very important for all fields in which cell cryopreservation is employed, particularly for clinical and multicenter studies wherein the comparability and reproducibility of results play a crucial role. To obtain reliable results and to maintain the quality of the cells, not only the freezing and thawing processes but also the storage conditions should be controlled and standardized, and any deviations should be documented.
机译：冷冻保存诸如细胞，组织和器官等生物材料是一个非常重要的普遍主题。它不仅用于许多研究领域，而且还用于临床领域。冷冻保存对于生殖医学和临床研究以及疫苗的开发非常重要。外周血单核细胞（PBMC）通常用于疫苗研究，其中不同研究机构和实验室之间的可比且可靠的结果非常重要。尽管对冷冻和解冻过程进行了充分的研究，控制和标准化，但通常不考虑储存条件。为了缩小这一差距，我们研究了低温存储过程中次优存储条件对PBMC生存能力，恢复能力和T细胞功能的影响。为此，在机器人系统的帮助下，将PBMC隔离并暴露在低温环境中（从0到350个温度波动周期，以50个周期为步长），以模拟大型生物存储库中的存储条件，并具有样品存储，移除和分类功能。模拟后，分析了生存力，恢复能力和T细胞功能，以确定温度升高的次数，这最终导致明显的细胞损伤。所有研究的参数随着温度循环次数的增加而降低。有时仅经过50个温度循环，就可以观察到明显的效果。这些结果对于所有应用细胞冷冻保存的领域都非常重要，特别是对于其中结果的可比性和再现性起关键作用的临床和多中心研究。为了获得可靠的结果并保持细胞的质量，不仅应控制和标准化冷冻和解冻过程，还应控制储存条件，并应记录任何偏差。

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