EFFECT OF DMSO-FREE CRYOPRESERVATION ON MSCS OSTEOGENESIS IN 3D POROUS SCAFFOLDS

摘要

Successful clinical application of bone tissue engineered constructs depends on efficiency of osteogenic differentiation of cells within 3D scaffolds. Dimethyl sulfoxide (DMSO)-free cryopreservation can have a direct effect on osteogenesis. Therefore, the aim of this research was to evaluate the ability of cryopreserved mesenchymal stromal cells (MSCs) to differentiate into osteogenic direction within 3D scaffolds.rnHuman adipose tissue-derived MSCs were cryopreserved without DMSO and serum using sucrose-based pretreatment protocol. After thawing non-cryopreserved and cryopreserved MSCs were seeded into gelatin, collagen and collagen / hydroxyapatite scaffolds, afterwards cell attachment, viability, metabolic and proliferation activity were tested. Efficiency of induced osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity and calcium content.rnAfter seeding into all investigated scaffolds cryopreserved and non-cryopreserved cells attached to their surface and were able to proliferate. Cryopreservation resulted in an increased ALP expression in all investigated groups; however had no effect on calcium contents. The maximum levels of ALP activity and calcium deposition were obtained for MSCs grown within collagen-based scaffolds, while cells within gelatin scaffolds demonstrated less efficiency of osteogenic differentiation.rnTo sum up, the developed DMSO- and fetal serum (FS) - free cryopreservation protocol preserves the potential of MSCs to differentiate towards osteogenic direction in 3D environment.