Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model.
Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model.
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DOI：
10.3390/cells11020278
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发表时间：
2022-01-14
期刊：
影响因子：
6
通讯作者：
Baust JG
中科院分区：
文献类型：
--
作者：
Baust JM;Snyder KK;Van Buskirk RG;Baust JG
在临床和发现科学中，基于细胞的复杂产品的开发和使用继续以前所未有的速度增长。为此，低温保存起着关键作用，作为一种使能过程，提供对生物材料的按需获取，促进活材料的大规模生产、储存和分配。尽管在过去的几十年中发挥了关键作用并取得了实质性的进步，但冷冻保存通常仍然是影响许多领域的瓶颈，包括细胞治疗，组织工程和组织库。研究已经说明了通过各种“前端”策略（如专用培养基、新的冷冻保护剂和冷冻保存期间的分子控制）控制冷冻保存诱导的延迟性细胞死亡（CIDOCD）的影响和益处。虽然证明是非常成功的，但大量的细胞死亡和细胞功能丧失仍然与冷冻保存有关。最近，我们专注于开发技术（RevitalICE™），旨在通过在解冻后恢复阶段缓冲细胞应激反应来减少CIDOCD的影响，以提高先前冷冻保存样品的回收率。在这项研究中，我们研究了在解冻后最初24小时内调节凋亡半胱天冬酶激活、氧化应激、未折叠蛋白反应和自由基损伤对总体细胞存活率的影响。将在传统的细胞外型和细胞内型冷冻保存培养基中体外冷冻保存的人造血祖细胞用作模型细胞系统以评估对存活的影响。我们的研究结果表明，通过调节这些途径中的几种，无论使用的冷冻培养基和二甲亚砜浓度如何，都可以改善细胞回收率。具体而言，通过使用氧化应激抑制剂，观察到总体活力平均增加20%。此外，结果表明，通过对在细胞内型培养基（Unisol™）中冷冻保存的样品使用解冻后回收试剂，获得了接近非冷冻对照的80%的总细胞存活率的改善。虽然总体生存率有所提高，但对特定细胞亚群和功能的影响的评估仍有待完成。虽然工作仍然存在，但这些结果代表了在开发用于发现科学以及商业和临床环境的改进的冷冻保存过程中迈出的重要一步。
The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, storage, and distribution of living materials. Despite serving a critical role and substantial improvements over the last several decades, cryopreservation often remains a bottleneck impacting numerous areas including cell therapy, tissue engineering, and tissue banking. Studies have illustrated the impact and benefit of controlling cryopreservation-induced delayed-onset cell death (CIDOCD) through various “front end” strategies, such as specialized media, new cryoprotective agents, and molecular control during cryopreservation. While proving highly successful, a substantial level of cell death and loss of cell function remains associated with cryopreservation. Recently, we focused on developing technologies (RevitalICE™) designed to reduce the impact of CIDOCD through buffering the cell stress response during the post-thaw recovery phase in an effort to improve the recovery of previously cryopreserved samples. In this study, we investigated the impact of modulating apoptotic caspase activation, oxidative stress, unfolded protein response, and free radical damage in the initial 24 h post-thaw on overall cell survival. Human hematopoietic progenitor cells in vitro cryopreserved in both traditional extracellular-type and intracellular-type cryopreservation freeze media were utilized as a model cell system to assess impact on survival. Our findings demonstrated that through the modulation of several of these pathways, improvements in cell recovery were obtained, regardless of the freeze media and dimethyl sulfoxide concentration utilized. Specifically, through the use of oxidative stress inhibitors, an average increase of 20% in overall viability was observed. Furthermore, the results demonstrated that by using the post-thaw recovery reagent on samples cryopreserved in intracellular-type media (Unisol™), improvements in overall cell survival approaching 80% of non-frozen controls were attained. While improvements in overall survival were obtained, an assessment on the impact of specific cell subpopulations and functionality remains to be completed. While work remains, these results represent an important step forward in the development of improved cryopreservation processes for use in discovery science, and commercial and clinical settings.
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影响因子：
1.6
作者：
Bissoyi, Akalabya;Pramanik, Krishna
通讯作者：
Pramanik, Krishna
影响因子：
8.8
作者：
Anderson, CD;Belous, A;Chari, RS
通讯作者：
Chari, RS
DOI：
10.1007/978-3-319-20579-3_4
发表时间：
2015-01-01
期刊：
BIOBANKING IN THE 21ST CENTURY
影响因子：
--
作者：
Baust, John M.;Corwin, William L.;Baust, John G.
通讯作者：
Baust, John G.
影响因子：
--
作者：
Baust, John M.;Vogel, Martin J.;Baust, John G.
通讯作者：
Baust, John G.
影响因子：
2.7
作者：
Baicu, SC;Taylor, MJ
通讯作者：
Taylor, MJ