Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function
Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function
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DOI：
10.1016/j.cryobiol.2013.06.012
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发表时间：
2013-10-01
期刊：
影响因子：
2.7
通讯作者：
von Briesen, Hagen
中科院分区：
文献类型：
--
作者：
Germann, Anja;Oh, Young-Joo;von Briesen, Hagen
分析来自生物库的冷冻保存的外周血单核细胞（PBMC）的抗原特异性免疫的能力对于评价对基于免疫的治疗的应答是必要的。为了确保具有可比性的检测结果，多中心试验中的合作研究需要可靠且可重复的冷冻保存，以保持细胞活力和功能。标准化的冷冻保存程序不仅包括样品收集、制备和冷冻，还包括在液氮中低温储存，没有任何温度波动，以避免细胞损伤。因此，我们开发了一种储存方法，以最大限度地减少次优储存条件，以最大限度地提高细胞活力，恢复和T细胞功能。我们比较了重复温度波动对细胞健康的影响，从样品储存，样品分选和去除相比，样品储存没有温度上升。我们发现，在低温储存期间的周期性温度变化降低了细胞活力、恢复和针对特异性抗原的免疫应答。我们表明，在保护罩系统下处理的样品，以避免或尽量减少这种重复的温度上升，具有可比的细胞活力和细胞回收率的样品储存没有任何温度波动。此外，T细胞的功能可以大大增加与使用的保护罩系统相比，没有这样的保护system.This数据表明，细胞完整性的温度波动的影响，应仔细考虑在未来的临床疫苗试验，并考虑最佳的样品储存条件。(C)2013作者爱思唯尔公司出版All rights reserved.
The ability to analyze cryopreserved peripheral blood mononuclear cell (PBMC) from biobanks for antigen-specific immunity is necessary to evaluate response to immune-based therapies. To ensure comparable assay results, collaborative research in multicenter trials needs reliable and reproducible cryopreservation that maintains cell viability and functionality. A standardized cryopreservation procedure is comprised of not only sample collection, preparation and freezing but also low temperature storage in liquid nitrogen without any temperature fluctuations, to avoid cell damage. Therefore, we have developed a storage approach to minimize suboptimal storage conditions in order to maximize cell viability, recovery and T-cell functionality.We compared the influence of repeated temperature fluctuations on cell health from sample storage, sample sorting and removal in comparison to sample storage without temperature rises. We found that cyclical temperature shifts during low temperature storage reduce cell viability, recovery and immune response against specific-antigens. We showed that samples handled under a protective hood system, to avoid or minimize such repeated temperature rises, have comparable cell viability and cell recovery rates to samples stored without any temperature fluctuations. Also T-cell functionality could be considerably increased with the use of the protective hood system compared to sample handling without such a protection system.This data suggests that the impact of temperature fluctuation on cell integrity should be carefully considered in future clinical vaccine trials and consideration should be given to optimal sample storage conditions. (C) 2013 The Authors. Published by Elsevier Inc. All rights reserved.