Abstract
Cellular cryopreservation is a platform technology which underpins cell biology, biochemistry, biomaterials, diagnostics, and the cold chain for emerging cell-based therapies. This technique relies on effective methods for banking and shipping to avoid the need for continuous cell culture. The most common method to achieve cryopreservation is to use large volumes of organic solvent cryoprotective agents which can promote either a vitreous (ice free) phase or dehydrate and protect the cells. These methods are very successful but are not perfect: not all cell types can be cryopreserved and recovered, and the cells do not always retain their phenotype and function post-thaw. This Perspective will introduce polyampholytes as emerging macromolecular cryoprotective agents and demonstrate they have the potential to impact a range of fields from cell-based therapies to basic cell biology and may be able to improve, or replace, current solvent-based cryoprotective agents. Polyampholytes have been shown to be remarkable (mammalian cell) cryopreservation enhancers, but their mechanism of action is unclear, which may include membrane protection, solvent replacement, or a yet unknown protective mechanism, but it seems the modulation of ice growth (recrystallization) may only play a minor role in their function, unlike other macromolecular cryoprotectants. This Perspective will discuss their synthesis and summarize the state-of-the-art, including hypotheses of how they function, to introduce this exciting area of biomacromolecular science.
Highlights
For the cryopreservation of mammalian cells, the standard protocol is freezing in a solution containing 5−10 wt % of the cryoprotective agent (CPA) dimethyl sulfoxide (DMSO), which was introduced in 19591 and remains the gold standard
DMSO causes differentiation of human leukemic cell lines,[5] as well as certain cells such as leukocytes and RAW 264.7 cells, which are highly sensitive to DMSO, with concentrations even below 1% significantly affecting ROS production.[6]
In addition to these specific instances, there is a concern with using DMSO for all cell lines due to its cytotoxicity at high concentrations and/or at room temperature.[7,8]
Summary
For the cryopreservation of mammalian cells, the standard protocol is freezing in a solution containing 5−10 wt % of the cryoprotective agent (CPA) dimethyl sulfoxide (DMSO), which was introduced in 19591 and remains the gold standard.
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