Cell, Tissue And Organ Preservation With Insect-Derived Antifreeze Peptides

We describe our efforts to date using Dendroides- and, to a lesser extent, Cujucus-derived recombinant antifreeze proteins for short- and long-term mammalian cell, tissue, and organ storage strategies. Some single Dendroides antifreeze proteins (DAFPs), DAFP-6 and DAFP-2 concentrations in combination with dimethylsulfoxide increased post-cryopreservation cell viability. However, no significant improvement in viability was observed with either DAFP-1 or DAFP-4. Furthermore, more complex DAFP formulations resulted in loss of viability enhancement observed with DAFP-6 and 2. Experiments were also performed to see if the combination of DAFPs-1, -2, and -4 plus thaumatin-like protein had a benefit for hypothermic storage at −7 versus +4 °C. Thaumatin-like proteins are polypeptides of about 200 residues originally described in plants that are synthesized in response to fungal infection. Wang and Duman (Biochemistry 45:1278–1284, 2006) had previously described a thaumatin-like protein from larvae of the beetle Dendroides canadensis that enhances the activity of AFPs.