Registration Number
0226U004699,
(0121U113329)
, R & D reports
Title
Determination of pre-treatment role for increasing the efficiency of cryopreservation and hypothermic storage of cellular structures with different levels of organization.
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Вивчити ефективність підходу попередньої обробки клітин сахарозою за умов моношарового культивування для зниження концентрації кріопротектора ДМСО та ксеногенних компонентів при кріоконсервуванні МСК з різних джерел. Дослідити вплив попередньої обробки сахарозою на життєздатність, метаболічну активність та диференціювальний потенціал МСК після кріоконсервування в складі тривимірних структур.
Head
Petrenko Oleksandr Yu., д.б.н.
Registration Date
06-11-2026
Organization
Institute of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
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To determine the impact of certain pre-treatment approaches on the efficiency of cryopreservation, hypothermic storage and experimental transplantation of cell structures with different levels of organization.
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The study focused on mesenchymal stem/stromal cells (MSCs) derived from human skin dermis, adipose tissue, and dental pulp, in both suspension and macroporous scaffold formats. The aim of the study was to evaluate the effectiveness of sucrose pretreatment as an approach to cryopreserving MSCs from different tissue sources and to investigate the effects of this pretreatment on MSC viability, metabolic activity, and differentiation potential following cryopreservation within three-dimensional structures. The results demonstrated that the optimal pretreatment conditions depend on the cellular source. Following cryopreservation in the absence of cryoprotectants and fetal bovine serum (FBS), the highest recovery of dermal MSCs was achieved after pretreatment with higher sucrose concentrations than those required for dental pulp- and adipose tissue-derived MSCs. Human MSCs isolated from the dermis, adipose tissue, and dental pulp, following sucrose pretreatment and cryopreservation in the presence of 5% dimethyl sulfoxide (DMSO) and in the absence of FBS, exhibited high recovery rates (over 80%). Upon return to culture conditions, the cells retained their ability to proliferate and undergo multilineage differentiation. It was established that cryopreservation of MSCs within three-dimensional matrices using controlled-rate freezing with 10% DMSO as a cryoprotectant preserves cell viability and metabolic activity at levels of 48% ± 4% and 40% ± 5%, respectively. The use of sucrose during pretreatment and subsequent cryopreservation with 10% DMSO as a cryoprotectant improved cell recovery within macroporous collagen scaffolds by 16%. Following cryopreservation, MSCs maintained in three-dimensional culture exhibited a typical fibroblast-like morphology, retained their multilineage differentiation potential, proliferated actively, and populated the entire volume of the collagen matrix.
Product Description
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Olena Y. Rohulska
Oleksandr M. Sukhach
Olena B. Revenko
Olha V. Ochenashko
Olha A. Semenchenko
Svitlana P. Mazur
Svitlana O. Vsevolodska
Natalia A. Trufanova
Daria V. Cherkashyna
Volodymyr Y. Prokopyuk
Oleksandra S. Hubenia
Volodymyr V. Zeienko
Ruslana A. Oberemok
Oleksii S. Mishchenko
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2026-11-06
