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Cell-compatible isotonic freezing media enabled by thermo-responsive osmolyte-adsorption/exclusion polymer matrices
Yui Kato1, Yuya Matsuda1, Takuya Uto2
1Faculty of Biological Science and Technology, Institute of Science and Engineering, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan.
Communications Chemistry
|November 29, 2023
Summary
New poly(zwitterion) cryoprotectants form cell-protecting matrices, enabling cryopreservation under isotonic conditions. This novel approach minimizes cell damage during long-term storage by controlling osmotic pressure.
Area of Science:
- Biotechnology
- Materials Science
- Cell Biology
Background:
- Cryopreservation requires cryoprotectants to prevent ice crystal formation.
- Conventional non-cell-permeable cryoprotectants cause cell damage due to high osmotic pressure and dehydration.
- Emerging polymer-type cryoprotectants form protective matrices but still require high osmotic pressure.
Purpose of the Study:
- To design a novel zwitterionic polymer (polyZI) for effective cryopreservation.
- To achieve cryopreservation of freeze-vulnerable cells under isotonic conditions.
- To investigate the mechanism of polyZI matrix formation and its effect on osmotic pressure.
Main Methods:
- Design and synthesis of a poly(zwitterion) (polyZI) with high cell membrane affinity.
- Application of polyZI as a non-cell-permeable cryoprotectant for freeze-vulnerable cells.
- Analysis of polyZI matrix formation, osmotic pressure regulation, and cell proliferation post-cryopreservation.
Main Results:
- PolyZI formed firm matrices around cells, enabling successful cryopreservation under isotonic conditions.
- The polyZI matrices regulated osmotic pressure by adsorbing/desorbing NaCl with temperature changes.
- Cell proliferation was initially delayed but recovered after washing with sucrose solution.
Conclusions:
- Poly(zwitterion)s can serve as effective non-cell-permeable cryoprotectants under isotonic conditions.
- The designed polyZI matrices offer a promising strategy to mitigate cryoinjury by controlling osmotic stress.
- Further optimization, including post-thaw washing, can improve cell recovery and proliferation.

