Gryshkov Oleksandr, Hofmann Nicola, Lauterboeck Lothar, Pogozhykh Denys, Mueller Thomas, Glasmacher Birgit
Institute for Multiphase Processes, Leibniz Universitaet Hannover, Callinstrasse 36, 30167 Hannover, Germany.
Institute for Transfusion Medicine, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Cryobiology. 2015 Aug;71(1):103-11. doi: 10.1016/j.cryobiol.2015.05.001. Epub 2015 May 14.
Multipotent stromal cells derived from the common marmoset monkey Callithrix jacchus (cjMSCs) possess high phylogenetic similarity to humans, with a great potential for preclinical studies in the field of regenerative medicine. Safe and effective long-term storage of cells is of great significance to clinical and research applications. Encapsulation of such cell types within alginate beads that can mimic an extra-cellular matrix and provide a supportive environment for cells during cryopreservation, has several advantages over freezing of cells in suspension. In this study we have analysed the effect of dimethyl sulfoxide (Me2SO, 2.5-10%, v/v) and pre-freeze loading time of alginate encapsulated cjMSCs in Me2SO (0-45 min) on the viability and metabolic activity of the cells after freezing using a slow cooling rate (-1°C/min). It was found that these parameters affect the stability and homogeneity of alginate beads after thawing. Moreover, the cjMSCs can be frozen in alginate beads with lower Me2SO concentration of 7.5% after 30 min of loading, while retaining high cryopreservation outcome. We demonstrated the maximum viability, membrane integrity and metabolic activity of the cells under optimized, less cytotoxic conditions. The results of this study are another step forward towards the application of cryopreservation for the long-term storage and subsequent applications of transplants in cell-based therapies.
源自普通狨猴（Callithrix jacchus，cjMSCs）的多能基质细胞与人类具有高度的系统发育相似性，在再生医学领域具有巨大的临床前研究潜力。细胞的安全有效长期储存对临床和研究应用具有重要意义。将此类细胞封装在藻酸盐珠内，藻酸盐珠可模拟细胞外基质并在冷冻保存期间为细胞提供支持性环境，这比将细胞悬浮冷冻具有多个优势。在本研究中，我们分析了二甲基亚砜（Me2SO，2.5 - 10%，v/v）以及藻酸盐封装的cjMSCs在Me2SO中的预冻加载时间（0 - 45分钟）对使用慢速冷却速率（-1°C/分钟）冷冻后细胞活力和代谢活性的影响。结果发现，这些参数会影响解冻后藻酸盐珠的稳定性和均匀性。此外，加载30分钟后，cjMSCs可在较低Me2SO浓度（7.5%）的藻酸盐珠中冷冻，同时保持较高的冷冻保存效果。我们在优化的、细胞毒性较小的条件下证明了细胞的最大活力、膜完整性和代谢活性。本研究结果朝着将冷冻保存应用于基于细胞疗法的移植长期储存及后续应用又迈进了一步。
