Determination of the membrane hydraulic permeability of MSCs

Abstract

A successful cryopreservation is based on knowledge of the optimal cooling rate. So far, this is often determined by way of complex parameter studies. Alternatively, the identification of cell specific characteristics, such as osmotic behaviour, membrane hydraulic permeability and activation energy could be used to calculate the optimal cooling rate. These parameters should be determined for supra-zero and sub-zero temperatures. In this study cryomicroscopy was used. Mesenchymal stromal cells (MSCs) from bone marrow were analysed. The determined membrane hydraulic permeability for sub-zero temperatures is significantly lower than that for supra-zero temperatures. On the contrary the activation energy is significantly higher in the presence of ice. The addition of a cryoprotective agent (CPA) such as dimethyl sulfoxid (DMSO) shows an additional influence on the characteristics of the membrane of the cell. The optimal cooling rate was determined with these parameters. For cryopreservation without DMSO the optimal cooling rate was found to be 12.82 K/min. If the MSCs were frozen with 5% (v/v) DMSO the optimal cooling rate is 16.25 K/min.

Keywords

• cryopreservation
• optimal cooling rate
• membrane hydraulic permeability
• mesenchymal stromal cells