The research on loading procedure for vitrification of NSCs

摘要

Vitrification is an "ice free" method of cryopreservation, which is reported to be capable of preserving stem cells in early passages and is of great potential for the clinic application. However, the osmotic damage and toxic damage due to the high concentration of cryoprotetant which is necessary for achieving vitrification should be considered, as the cells would dehydrate acutely when the difference of osmotic pressure between intra- and extra- cellular is large enough. This dehydration would lead to cell shrinkage and even death. In order to decrease these damages, different loading methods were designed, and a hypothesis was proposed that the more cell volume shrunk, and the longer cells in touch with the high concentration solution, the more damage was produced. The viability after removing cryoprotectant was determined by CCK-8 kit. As a result, the multistep loading procedure with gradually reduced time intervals showed the least change of cell volume and the best viability.