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Registration Number

0222U005142, 0122U200072 , R & D reports

Title

Cryobiology profile of 3D mesenchymal stem cells spheroids

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Head

Petrenko Oleksandr Yu., д.б.н.

Registration Date

20-12-2022

Organization

Institute of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine

Description

 The aim of the work is to preserve viability, metabolic and secretory activity, the ability to proliferate and multilineage differentiation of MSCs in spheroids using cryobiological approaches - cryopreservation and hypothermic storage. Methods of monolayer and three-dimensional cell cultivation, light and fluorescence microscopy, multicomponent immunoassay, cryopreservation, and hypothermic storage were used. It is shown that slow-rate two-stage freezing according to the standard protocol under the protection of 10% DMSO leads to partial disruption of spheroids integrity and loss of a significant part of viable cells in their composition. Cryopreservation according to a modified protocol with a reduced concentration of DMSO (7.5%) and addition of sucrose or Rho-kinase inhibitors is more effective and allows obtaining functionally complete cells. The effect of hypothermic storage on the metabolic activity of MSCs in spheroids was established. It was shown that the original preservation solutions - sucrose and trehalose containing - ensure high rate of cell preservation in spheroids after 7 days of storage at temperature of 4°С. After a week of hypothermic storage, MSCs remain morphologically, metabolically and functionally complete: they can adhere, proliferate, unergo induced differentiation and have high secretory activity.

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Mazur Svitlana P.
Revenko Olena B.
Rogulska Olena Yu.
Tykhvynska Olga O.
Trufanova Natalia A.