Biopolym. Cell. 2024; 40(1):37-46.
Molecular and Cell Biotechnologies
Study of physical processes occurring in serum-containing and polymer-based serum-free cryoprotective media
1Pakhomov O. V., 1Gurina T. M., 1Polyakova A. L., 2Mazaeva V. S., 3Deng B., 1Bozhok G. A.
  1. Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine
    23, Pereyaslavskaya Str., Kharkiv, Ukraine, 61016
  2. Department of Studies of Technology for Processing Oils and Fats, Ukrainian Research Institute of Oils and Fats, NAAS of Ukraine
    2a, Dziuby Ave., Kharkiv, Ukraine, 61019
  3. Department of Physiology, Henan University of Science and Technology
    263, Kaiyuan Ave., Henan, Luoyang, China, 471023


Aim. To investigate the mechanisms of cryoprotection with compositions that were supplemented with polymers (dextran [Dex], hydroxyethyl starch [HES], polyethylene glycol [PEG]) or fetal bovine serum [FBS]). Methods. Differential scanning calorimetry (DSC) and thermomechanical analysis (TMA) of cryoprotective media containing serum, as well as media that do not contain serum or its components but include polymers. Results. The combinations of dimethyl sulfoxide with FBS or polymers in cryoprotective media were able to promote the formation of amorphous phase and suppress salt eutectic crystallization/melting and recrystallization.The medium supplemented with 0.7 M Me2SO and 100 mg/ml Dex (0.7Me2SO+Dex) was somewhat superior compared to other compositions. It also had an optimal glass transition temperature (-26…-21°C). The feature correlated with an adequate cell recovery when the medium was utilized to preserve testicular cells. Conclusions. The mechanism of cryoprotection depends not only on the concentrations of cryoprotective agents such as Me2SO, polymers or FBS, but also on the composition of the cryoprotective media. The 0.7Me2SO+Dex medium can be the basis for the creation of commercial serum-free cryoprotective media with the defined stable composition that does not require the removal of Me2SO before using the cryopreserved cells or tissues for cultivation, transplantation and transfusion.
Keywords: serum-/xeno-free media, dextran, hydroxyethyl starch, polyethylene glycol, differential scanning calorimetry, cryopreservation


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