Rogulska O. Cryopreservation, monolayer and three-dimensional culture of multipotent mesenchymal stromal cells

The research object was the realization of proliferative and differentiation potential of human multipotent mesenchymal stromal cells (MSCs) in monolayer and three-dimensional culture conditions and after cryopreservation in serum-free media with platelet lysate and saccharides. The research aim ? determination of effect of expansion and cryopreservation conditions on morphological and functional characteristics of MSCs in monolayer and three-dimensional culture within matrices of different nature. Research methods: in vitro cell culture, programmable freezing, cytochemistry, spectrophotometry, fluorescence spectroscopy, light, fluorescent and scanning electron microscopy, morphometric analysis, statistical analysis of the results. For the first time the efficiency of the saccharides (sucrose, trehalose, raffinose) application for pretreatment and following cryopreservation of MSCs in the absence of dimethylsulphoxide and the embryonic serum was shown. It was established that platelet lysate presence in the culture medium compared with ES promoted proliferation and colony-forming potential of MSCs and enhanced efficacy of osteogenic differentiation. For the first time it was proven that PL application at different stages of cryopreservation improved safety, viability and cell survival. For the first time, the comparative analysis of the adhesion, proliferation and differentiation of non-cryopreserved and cryopreserved MSCs when cultured in natural and artificial three-dimensional matrices based on chitin, chitosan, collagen, collagen with hydroxyapatite inclusions, gelatin and alginate was carried out.