Serum-free non-toxic freezing solution for cryopreservation of human adipose tissue-derived mesenchymal stem cells

Abstract

To develop a cost-effective, non-toxic and xeno-free freezing solution for the preservation of adipose tissue-derived stem cells (hADSC) with a long shelf-life. The potential of various hydrocolloids and organic osmolytes as cryoprotectants and individual components of phosphate buffered saline (PBS) as carrier media were evaluated to formulate a freezing solution for the cryopreservation of hADSCs. Among the hydrocolloids, the highest viability, 55%, was achieved with post-thawed (after 48h storage at -80°C) hADSCs cryopreserved in 10% (v/v) polyvinylpyrrolidone (PVP) using PBS as carrier media. 0.9% NaCl was a superior carrier medium resulting an enhanced cell viability (70%) when used in 10% PVP than other components of PBS. A higher cell viability (81%) was achieved when 10% PVP/0.9% NaCl was supplemented with 60mM ectoin. The cryopreserved cells retained normal cytoskeletal distribution pattern and adipogenic and osteogenic differentiation ability during 14 and 21days of incubation. A serum-free and non-toxic 10% PVP/0.9% NaCl/60mM ectoin freezing solution was developed for cryopreservation of hADSC for application in tissue engineering and regenerative medicine.