Gorokhova N. Cryopreservation of cultured fibroblast-like cells by slow cooling and vitrification

Dissertation is aimed to investigate the morphologycal and functional state of cultured human fibroblast-like cells (FLC) of early organogenetic stage after cryopreservation by slow cooling under protection of dimethyl sulfoxide (DMSO) as well as by vitrification in multicomponent cryoprotective solutions. It was obtained that about 95% of 4 - 12 passages FLC express immunophenotype of mesenchymal stromal cells: positive for CD29, CD44, CD73, CD105 and negative for CD34, CD38, CD45. Cultured FLC possess the ability to differentiate into osteogenic and adipogenic lineages. Cryopreservation under protection of DMSO in final concentration of 10% using cooling rate 1 C per min allow to preserve in a great extent the viability, metabolic activity and induced osteogenic and adipogenic differentiation capacity of FLC. The possibility of effective cryopresevation of FLC suspension by vitrification was studied. The application of solution which contained DMSO, ethylene glycol, 1,2-propanediol and sucrose for vitrification of cultured FLC allow to save their morphologycal and functional characteristics as well as ability to differentiate into osteogenic and adipogenic lineages