Development of novel cryoprotectants and their application for tissue engineering

Abstract

Cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO), glycerol, or ethylene glycol have been used for the cryopreservation of many types of cells. DMSO is the most effective CPA but shows cytotoxicity and can effect differentiation. Here we report that the novel polymeric CPAs having low toxicity and high efficiency for mammalian cells including stem cells. Carboxylated poly-L-lysine (COOH-PLL) showed higher cryopreservation efficiency for many types of cell than the conventional CPAs. In addition, mesenchymal stem cell monolayers were cryopreserved more effectively than with DMSO based cryoprotective solutions via slow or rapid freezing methods. These results indicate that these polymeric extracellular CPAs may replace current CPAs and the high viability after thawing and nonnecessity of serum ensure that these CPAs may be used in tissue engineering fields for the storage of tissue constructs.