Ice Recrystallization Inhibitors enable efficient cryopreservation of induced pluripotent stem cells: a functional and transcriptomic analysis

Kathleen Mommaerts, Satoshi Okawa, Margaux Schmitt, Olga Kofanova, Tracey Turner, Robert Ben, Antonio Del Sol, Jens C. Schwamborn, Jason P. Acker, Fay Betsou


The successful use of human induced pluripotent stem cells (iPSCs) for research or clinical applications requires the development of robust, efficient and reproducible cryopreservation protocols. After cryopreservation, the survival rate of iPSCs is poor and cell line dependent. The aim of this study was to assess the use of Ice Recrystallization Inhibitors (IRIs) for cryopreservation of human iPSCs. A toxicity screening study was first performed to select the specific small-molecule IRI and concentration for further evaluation. Then, a cryopreservation study compared the cryoprotective efficiency of 15 mM IRIs in 5% or 10% DMSO-containing solution with CryoStor® CS10. Three iPSC lines were cryopreserved as single-cell suspension in cryopreservation solutions and post-thaw characteristics including pluripotency and differential gene expression were assessed. We demonstrate the fitness-for-purpose of 15 mM IRI in 5% DMSO as an efficient cryoprotective solution for iPSCs in terms of their post-thaw recovery, viability, pluripotency and transcriptomic changes. Given that this dataset is the first one where mRNA sequencing has been used to identify expression changes resulting from iPSCs cryopreservation, it has the potential to be used for molecular mechanism analysis relating to cryopreservation. IRIs can reduce DMSO concentrations, thereby improving the utility, effectiveness and efficiency of cryopreservation.


The data is available here.


The scripts used to analyse the data are available here.