Lauterboeck L, Hofmann N, Mueller T, Glasmacher B
Institute for Multiphase Processes, Leibniz Universität Hannover, Hannover, Germany.
Service Unit Embryonic Stem Cells, Institute for Transfusion Medicine, Medical School Hannover, Germany.
Cryobiology. 2015 Dec;71(3):384-90. doi: 10.1016/j.cryobiol.2015.10.145. Epub 2015 Oct 21.
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 °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.
低温保存是一种在再生医学中被广泛用于储存多能间充质基质细胞（MSCs）的技术。因此，在提高细胞活力和功能方面改进当前的低温保存程序很重要。在本研究中，我们优化了普通狨猴（Callithrix jacchus，cj）来源的MSCs的低温保存方案，普通狨猴可作为各种病理和移植研究中的非人灵长类动物模型，在再生医学中具有巨大潜力。我们研究了在广泛的温度范围内通过诱导成核来主动控制成核温度以及两种不同二甲基亚砜浓度（Me2SO，5%（v/v）和10%（v/v））的效果，以评估对解冻后细胞活力、代谢活性和复苏的总体影响。当在-10°C诱导结冰时，存活率和代谢活性表现最佳。低温显微镜研究表明，不同成核温度下冰晶形态以及细胞内结冰情况存在差异。零下较高的成核温度导致较大的细胞外冰晶和细胞脱水，而零下较低的成核温度导致较小的冰晶和细胞内结冰。
