Novikova O. Morphofunctional features of cryopreserved neural crest derived cells obtained from different sources

The object of the study is the effect of cryopreservation in cryoprotectant media based on dimethyl sulfoxide (DMSO) and its combination with protein-peptide supplements on morphological, phenotypic, cytological features of 2D- and 3D-cultures obtained from adrenal glands (AG) and dermal papillae (DP) of neonatal animals. The goal was to study the effect of cryopreservation in a cryoprotectant medium based on DMSO and protein-peptide supplements on morphological, functional, cytogenetic properties of 2D- and 3D-cultures obtained from AG and DP of neonatal animals. Research methods: enzymatic isolation of cells from adrenal tissue, explants for isolation of DP vibrissae cell cultures, monolayer (2D) and three-dimensional (3D) cultivation, controlled-speed freezing; immunocytochemical, cytofluorimetric methods - to determine the expression of immunophenotypic markers, cytological analysis methods - to study the mitotic rate and pathologies of DP cell mitosis, light and fluorescence microscopy - to study cell morphology, morphometric analysis, methods of statistical analysis. It was found for the first time that cultivation in the serum-free medium promotes the spontaneous formation of multicellular spheroids (MS) in the culture of neonatal rabbit DP. It was found for the first time that in the cryoprotective media based on DMSO and after cryopreservation, a long-term effect is observed in the DP cell culture, which consists in the growth of pathological cell divisions and slowing down the growth of MS. Safe and effective concentrations of DMSO (5 - 7.5%) during cryopreservation of these objects have been established. For the first time, the effect of cryopreservation on the character of expression of chromogranin A - protein of secretory granules of chromaffin cells before and after cryopreservation of neonatal piglets AG cell culture was analyzed. The redistribution of subpopulations in the culture of AG cells in the cultivation process, which is manifested in a relative increase of the number of cells in the MS was established. Field of Application: cryobiology, biology, medicine, biotechnology, veterinary, pharmacy.