The use of an icebindingprotein out of the snowflea Hypogastrura harveyi as a cryoprotectant in the cryopreservation of mesenchymal stem cells

Today researchers look for a possibility to keep cells for a long time without losing their viability. For that cryopreservation is often used. In this process it is necessary that the cells are not destroyed so cryoprotective agents (CPA) are needed. At the moment 5 to 10 % dimethylsulfoxide (DMSO) is mostly used, but this chemical is cytotoxic to the cells. So an alternative is needed. In this work experiments are made with an icebindingprotein (IBP) of Hypogastrura harveyi, as an alternative to DMSO. It was shown in previous studies that this protein isn’t cytotoxic for the cells, with crude extract and purified inclusion bodies it even seems that the mixtures have a positive effect on growth and proliferation. As a first step the protein was produced heterologous in E. coli and then the crude extract and the purified inclusion bodies were used for experiments on the influence of the IBP on the cryopreservation of mesenchymal stem cells from the common marmoset monkey Callithrix jacchus. In the process it was found that the protein could not replace DMSO completely. But it was possible to show that the DMSO-concentration can be reduced by adding the IBP.