Active control of the nucleation temperature enhances freezing survival of multipotent mesenchymal stromal cells
Active control of the nucleation temperature enhances freezing survival of multipotent mesenchymal stromal cells
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DOI：
10.1016/j.cryobiol.2015.10.145
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发表时间：
2015-12-01
期刊：
影响因子：
2.7
通讯作者：
Glasmacher, B.
中科院分区：
文献类型：
--
作者：
Lauterboeck, L.;Hofmann, N.;Glasmacher, B.
超低温保存是一种广泛应用于多能间充质间质细胞（MSCs）储存的技术。因此，在提高细胞活力和功能方面，改进目前的冷冻保存程序是很重要的。在本研究中，我们优化了来自普通狨猴（Callithrix jacchus, cj）的间充质干细胞的冷冻保存方案，该物种可作为非人灵长类动物模型用于各种病理和移植研究，具有很大的再生医学潜力。在较宽的诱导成核温度和两种不同二甲亚砜浓度（Me2SO, 5% （v/v）和10% (v/v)）下，研究了主动控制成核温度对细胞存活率、代谢活性和解冻后恢复的总体影响。低温显微镜研究表明，不同成核温度下的冰晶形态和细胞内冰的形成存在差异。低温成核导致细胞外冰晶变大，细胞脱水；低温成核导致细胞内冰晶变小，形成细胞内冰。(C) 2015爱思唯尔公司版权所有。
Cryopreservation is a technique that has been extensively used for storage of multipotent mesenchymal stromal cells (MSCs) in regenerative medicine. Therefore, improving current cryopreservation procedures in terms of increasing cell viability and functionality is important. In this study, we optimized the cryopreservation protocol of MSCs derived from the common marmoset Callithrix jacchus (cj), which can be used as a non-human primate model in various pathological and transplantation studies and have a great potential for regenerative medicine. We have investigated the effect of the active control of the nucleation temperature using induced nucleation at a broad range of temperatures and two different dimethylsulfoxide concentrations (Me2SO, 5% (v/v) and 10%, (v/v)) to evaluate the overall effect on the viability, metabolic activity and recovery of cells after thawing. Survival rate and metabolic activity displayed an optimum when ice formation was induced at -10 degrees C. Cryomicroscopy studies indicated differences in ice crystal morphologies as well as differences in intracellular ice formation with different nucleation temperatures. High subzero nucleation temperatures resulted in larger extracellular ice crystals and cellular dehydration, whereas low subzero nucleation temperatures resulted in smaller ice crystals and intracellular ice formation. (C) 2015 Elsevier Inc. All rights reserved.