Development of a Cryoprotective Formula Based on the Fucose-Containing Polysaccharide FucoPol

This study reports the performance of the polysaccharide FucoPol as an enhancer of cryoprotective formulations. FucoPol at a concentration of 0.25% (w/v) was added to several normothermic Dulbecco-derived solutions and hypothermic Euro-Collins, Custodiol-HTK, and Unisol-CV media, substituting some constituents in the latter class to develop FucoPol-based formulations that were tested for their ability to cryopreserve Vero cells. Supplementation yielded post-thaw cell recovery enhancements of at least 70% and averaged at 82%. The FucoPol-supplemented formulations Dulbecco(+)+FP and Unisol-CVS3 achieved cell viabilities capable of competing with the commercial cryogenic formula CryoStor CS5. Particularly in Unisol-CVS3, mannitol, glucose, gluconate, and dextran were all substituted by 0.25% FucoPol, and still, a similar viability was achieved. Multiparametric correlation clustering showed that FucoPol cryoprotection synergizes best with K+, Ca2+, and Cl- in its microenvironment. Component substitution analysis demonstrated that FucoPol acts consistently as a cryoprotectant, an impermeant, and a colloidal stabilizer, providing a combined alternative to glucose, mannitol, gluconate, and dextran, thus highlighting its potential in the development of high-performing cryoprotective formulas.