Cryoprotectants for the vitrification of corneal endothelial cells
Fan WX., Ma XH., Ge D., Yu XC., Liu TQ., Cui ZF.
This work was to select and test the possible cryoprotective agents (CPAs) and to obtain a suitable formula for vitrification of corneal endothelial cells (CECs). Fresh bovine CECs were isolated and tested according to an optimized vitrification protocol with multi-step CPA loading and removal. Two types of CPA components, i.e. the penetrating CPA components and nonpenetrating saccharide CPAs were experimentally evaluated via using the viability measured by trypan blue. Dimethyl sulfoxide (Me2e glycol (EG), 1, 2-propanediol (1, 2-PD), 2, 3-butanediol (2, 3-BD), acetamide (Ace) and ethylene glycol monomethyl ether (EGMME) were chosen as the penetrating CPA components, and xylose, fructose, mannose, glucose, maltose, sucrose and trehalose were used as the saccharide CPAs, respectively. The results show that EG is the most suitable penetrating CPA component and glucose the best saccharide CPA. The optimized concentrations for each component in the chosen vitrification solution are 52% EG and 8% glucose. The CEC survival ratio of (85.5±0.7)% (mean±S.D.) was obtained following the established protocol.