DMSO-free preservation of cellular therapies and regenerative medicine products

Diverse biological systems (plants, insects, etc.) survive high salt environments, dehydration, drought, freezing temperatures and other stresses through the use of osmolytes. In the human kidney, a mixture of five osmolytes are used to stabilize the cells.  We hypothesized that, just as in nature, multicomponent osmolyte solutions (sugars, sugar alcohols and amino acids) can be used to preserve cells effectively. We have confirmed the hypothesis that combinations of sugars, sugar alcohols and amino acids can be effective in preserving cells used therapeutically. The focus of this project is to expand the number of cells that can be stabilized using osmolytes and translating the protocols developed into clinical and commercial application.  This includes the production of chemically defined preservation solutions with better and more consistent performance than "home-brew" solutions.

Multicellular systems (aggregates and tissues) exhibit increased sensitivity to freezing.  We propose to use strategies developed by nature to stabilize more complex organisms and enhance stability of multicellular systems.  This includes exploring Natural Deep Eutectic Systems (NADES) and multicomponent osmolyte solutions for cryogenic and high-subzero storage.  The addition of mechanistic preservation molecules, to target cell-specific mechanisms of damage, are being explored and compared to other cell types.  


Actin alignment of fresh mesenchymal stromal cells (A) compared to thawing from cells preserved in a DMSO-free solution (B) and DMSO based solution (C)

Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions - Kathryn Pollock, Rebekah M. Samsonraj, Amel Dudakovic, Roman Thaler, Aron Stumbras, David H. McKenna, Peter I. Dosa, Andre J. van Wijnen, and Allison Hubel


Immunocytochemistry (A,B) and karyotype (C) of human induced pluripotent stem cells (hiPSCs) cryopreserved using an optimized dimethyl sulfoxide (DMSO)-free solution


Cryopreservation of Human iPS Cell Aggregates in a DMSO-Free Solution—An Optimization and Comparative Study - Rui Li, Kathlyn Hornberger, James R. Dutton, and Allison Hubel