Intracellular delivery of trehalose renders mesenchymal stromal cells viable and immunomodulatory competent after cryopreservation
| Autor: | Janja Zupan, Miomir Knežević, Damijan Miklavčič, Ariana Barlič, Barbara Dovgan |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Stromal cell Clinical Biochemistry Cell Biomedical Engineering Mesenchymal stromal cells Bioengineering Cryopreservation Immunomodulation 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine medicine Extracellular Electroporation Mesenchymal stem cell Trehalose Cell Biology Cell biology 030104 developmental biology medicine.anatomical_structure chemistry 030220 oncology & carcinogenesis Original Article Intracellular Biotechnology |
| Zdroj: | Cytotechnology |
| ISSN: | 1573-0778 0920-9069 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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