Trehalose is a nontoxic disaccharide and a promising cryoprotection agent for medically applicable cells. In this study, the efficiency of combining trehalose with reversible electroporation for cryopreservation of two types of human mesenchymal stromal cells was investigated: adipose-derived stromal cells, and umbilical-cord-derived stromal cells. Comparable results to standard dimethyl sulfoxide cryopreservation protocols were achieved, even without extensive electroporation parameters and protocol optimization. The presence of high extracellular trehalose resulted in comparable cell viabilities without and with electroporation. According to the determination of trehalose concentrations, 250 mM extracellular trehalose resulting in, 20 mM to 50 mM intracellular trehalose were sufficient for successful cryopreservation of cells. With electroporation, higher (i.e. 50 mM to 90 mM) intracellular trehalose was achieved after cryopreservation, although cell survival was not improved significantly. To evaluate the impact of electroporation and cryopreservation on cells, stress and immune-activation-related gene expression were analyzed. Electroporation and/or cryopreservation resulted in increased SOD2 and HSPA1A expression. Despite the increased stress response, the high up-regulation by mesenchymal stromal cells of immunomodulatory genes in the inflammatory environment was not affected. Highest expression was seen for the IDO1 and TSG6 genes. In conclusion, cryopreservation of mesenchymal stromal cells in trehalose results in comparable characteristics to their cryopreservation using dimethyl sulfoxide.

海藻糖是一种无毒的二糖，是一种有前途的医学适用细胞的冷冻保护剂。在这项研究中，研究了海藻糖与可逆电穿孔相结合用于冷冻保存两种类型的人间充质基质细胞的效率：脂肪来源的基质细胞和脐带来源的基质细胞。即使没有广泛的电穿孔参数和协议优化，也获得了与标准二甲基亚砜冷冻保存协议相当的结果。高细胞外海藻糖的存在导致在没有和有电穿孔的情况下相当的细胞活力。根据海藻糖浓度的测定，250 mM 细胞外海藻糖产生，20 mM 至 50 mM 细胞内海藻糖足以成功冷冻保存细胞。使用电穿孔，冷冻保存后细胞内海藻糖含量较高（即 50 mM 至 90 mM），但细胞存活率并未显着提高。为了评估电穿孔和冷冻保存对细胞的影响，分析了应激和免疫激活相关基因的表达。电穿孔和/或冷冻保存导致增加SOD2和HSPA1A表达。尽管应激反应增加，炎症环境中免疫调节基因间充质基质细胞的高度上调并未受到影响。IDO1和TSG6基因的表达最高。总之，间充质基质细胞在海藻糖中的冷冻保存与使用二甲基亚砜冷冻保存的特征相当。