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Trehalose cryopreservation of human mesenchymal stem cells from cord and adipose tissue

R Aguilar-Quesada(1) N Izaguirre-Pérez(1) P A Aguilar-Solana(1) G Ligero(1) J A Carrillo-Ávila(1) I Biunno(2) P Catalina(1)

1:Coordinating Node, Andalusian Public Health System Biobank, Granada; 2:Institute of Genetic and Biomedical Research, National Research Council, Milan

Long-term cryopreservation of human mesenchymal stem cells (MSCs), for regenerative medicine applications, is highly important both in research and therapeutics. Current cryopreservation methods are based on dimethyl sulphoxide (DMSO) as cryoprotectant agents (CPAs), which has been shown to be toxic in clinical applications. Therefore, an alternative solution for cryopreservation of MSCs is required and we assessed the use of trehalose. The effects of trehalose as CPA at different concentrations, alone or in combination with ethylene glycol (EG) and glycerol (GLY)), was tested in MSCs from cord tissue through determination of cell viability, identity, proliferation and migration capacity. Although trehalose cryopreserved MSCs showed lower viability comparing with DMSO but cells maintained their functional properties, obtaining the best results when 0.5 M trehalose + 10% ethylene glycol (EG) was used to cryopreserve MSCs, also from adipose tissue.  Cells viability, recovery capacity, stability and stress-related gene expression for both, MSCs from cord and adipose tissue, after cryopreservation with 0.5 M trehalose + 10% ethylene glycol (EG) were analysed and results will be presented. In conclusion, preliminary assays show that trehalose-based solution could be considered an effective and less toxic alternative to DMSO for cryopreservation in liquid nitrogen of MSCs.

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