Effect of cryopreservation on viability and growth efficiency of stromal-epithelial cells derived from neonatal human thymus

• Published in: Cryobiology 2017-10, Vol.78, p.70-79
• Main Authors: Shichkin, Valentin P., Gorbach, Oleksandr I., Zuieva, Olga A., Grabchenko, Nataliia I., Aksyonova, Irina A., Todurov, Boris M.
• Format: Article
• Language: English
• Subjects: Cell phenotype; Cell Proliferation - drug effects; Cell Survival - drug effects; Cold Temperature; Cryopreservation; Cryopreservation - methods; Cryoprotective Agents - pharmacology; Dextrans - pharmacology; Dimethyl Sulfoxide - pharmacology; Epithelial Cells - cytology; Glycerol - pharmacology; Heart Diseases - congenital; Human thymus; Humans; Hydroxyethyl Starch Derivatives - pharmacology; Liquid nitrogen; Long-term storage; Stromal-epithelial cell cultures; Sucrose - pharmacology; Thymus Gland - cytology; Thymus Gland - embryology
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1016/j.cryobiol.2017.06.010

The thymus is the major site of T lymphocyte generation and so is critical for a functional adaptive immune system. Since, thymectomy is a component of neonatal surgery for congenital heart diseases, it provides great potential for collection and storage of thymic tissue for autologous transplantation. However, specific investigation into the optimum parameters for thymic tissue cryopreservation have not been conducted. In this research, we evaluated the effect of different cryoprotective media compositions, which included penetrating (Me2SO, glycerol) and non-penetrating (dextran-40, sucrose, hydroxyethyl starch) components, on the viability and functionality of frozen-thawed human thymic samples to select an optimal cryoprotective medium suitable for long-term storage of thymic tissue and a stromal-epithelial enriched population. Our primary focus was on receiving, low-temperature storage, culturing and evaluation of thymic tissue samples from newborns and infants with congenital heart diseases, who had undergone thymectomy as a part of standard surgical procedure. Thus, this work builds the platform for autologous clinical intervention into the thymus-deficient patients with congenital heart diseases. From our data, we conclude that although there were no significant differences in efficiency of tested cryoprotective media compositions, the combination of Me2SO and dextran-40 compounds was the most suitable for long-term storage both thymic cell suspensions and thymic fragments based on the viability of CD326+ epithelial cells and stromal-epithelial cell monolayer formation. [Display omitted] •Effect of different cryoprotectants on viability of human thymic samples.•Optimal cryoprotectant for long-term storage of thymic tissue and epithelial cells.•Monolayer formation by frozen-thawed thymic stromal-epithelial cells.