Osteogenic potential of multipotent mesenchymal stromal cells from human exfoliated deciduous teeth before and after cryopreservation

The use of multipotent mesenchymal stromal cellsfrom human exfoliated deciduous teeth (SHED) to stimulate bone regeneration requires data on the influence of cryopreservation on the osteogenic differentiation capacity of these cells. SHED were subjected to cryopreservation. Before freezing and after thawing, cell cultures were exposed osteogenic differentiation with vitamin D3 or dexametasone and assessed for expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 using real-Time qPCR. Extracellular matrix (ECM) mineralization was evaluated by Alizarin red staining. Supplementation of osteogenic medium with vitamin D3 increased the expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 as well as promoted an increase in the synthesis and mineralization of ECM in the cells both before and after cryopreservation. In the presence of vitamin D3 gene expression of alkaline phosphatase, BMP-2 and RunX2 after cryopreservation was higher than before freezing. Gene expression of osteocalcin, BMP2 RunX2 in osteogenic medium with vitamin D3was higher compared with dexamethasone for 14 days differentiation both before or after cryopreservation. The maintenance of SHED osteogenic differentiation potential after long-Term cryopreservation provides a basis for banking of these cellsfor further auto- or allotransplantation.