Abstract
Cryoprotective agents (CPAs) are typically solvents or small molecules, but there is a need for innovative CPAs to reduce toxicity and increase cell yield, for the banking and transport of cells. Here we use a photochemical high-throughput discovery platform to identify macromolecular cryoprotectants, as rational design approaches are currently limited by the lack of structure–property relationships. Using liquid handling systems, 120 unique polyampholytes were synthesized using photopolymerization with RAFT agents. Cryopreservation screening identified “hit” polymers and nonlinear trends between composition and function, highlighting the requirement for screening, with polymer aggregation being a key factor. The most active polymers reduced the volume of dimethyl sulfoxide (DMSO) required to cryopreserve a nucleated cell line, demonstrating the potential of this approach to identify materials for cell storage and transport.
Highlights
Cryoprotective agents (CPAs) are typically solvents or small molecules, but there is a need for innovative cryoprotective agent (CPA) to reduce toxicity and increase cell yield, for the banking and transport of cells
Current cryopreservation protocols for mammalian cells rely on the addition of high concentrations of dimethyl sulfoxide (DMSO) as the cryoprotective agent (CPA)
DMSO does not give full recovery of all cells post-thaw and is intrinsically cytotoxic.[5−7] DMSO does not protect against all mechanisms of cell death
Summary
Cryoprotective agents (CPAs) are typically solvents or small molecules, but there is a need for innovative CPAs to reduce toxicity and increase cell yield, for the banking and transport of cells. Using liquid-handling systems and photo-RAFT polymerization, a library of polymers were synthesized, characterized, and screened for cryopreservation.
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