Prokopiuk V.Yu.*, Shevchenko M.V.*, Musatova I.B. *, Skybina K.P. **, Prokopiuk O.S.* ,Kozub M.I. **
*Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
**Kharkiv Medical Academy of Postgraduate Education of Ministry of Health of Ukraine
Toufik’s Medical Journal
Volume 1, Supplement 1, November 2021
Abstract from Biomedical Perspectives III
Introduction: The development of cell therapy and experimental medicine requires the use of storage technologies for cells, tissues and bioengineered constructions. Mesenchimal stеm cells (MSCs) have shown great potential in the treatment of various diseases. However, despite a sufficient number of studies on the storage of MSCs at positive temperatures, the optimal conditions for short-term storage in subnormothermic and hypothermic conditions, allowing to obtain high indices of cell viability, have not been studied enough.
Aim: Search for optimal storage conditions for MSCs at positive temperatures with maximum preservation of their morphofunctional characteristics for use as a biomaterial in experimental medicine.
Materials and methods: MSCs from mice placentas (PMSCs) were isolated by enzymatic method, cultured at a concentration of 2×105/hole in a CO2 incubator at 37°C in an atmosphere of 5% CO2 for 10 days. For hypothermic (4°C, HS) or subnormothermic storage (20°C, SNS) of cells, DMEM culture medium enriched with 10% FBS was used. PMSCs were stored in 10 ml polystyrene tubes with a non-adherent surface at a concentration of 2×105/ml. The concentration of PMSCs in the suspension, their viability and cultural characteristics were assessed after 1, 2, 3 and 4 days of storage.
Results: The concentration of PMSCs in the suspension, their viability, adhesion ability and proliferative activity remained at the control level at SNS for 3 days. After 4 days of SNS, a slight decrease in the rate of monolayer formation in culture was observed. The level of PMSCs recovery after HS was lower: after 1 day of storage, there was a significant decrease in concentration of cells in the suspension, their viability, adhesion ability, and the ability to form a monolayer in culture. After 2 days of HS the cells lost their ability to proliferate and form a monolayer.
Conclusion: PMSCs, as a biomaterial for use in experimental medicine, can be stored in culture medium at a temperature of 20°C for up to 3 days while maintaining high indices of viability, concentration and cultural characteristics.