Nonionic and Water-Soluble Poly(<scp>d</scp>/<scp>l</scp>-serine) as a Promising Biomedical Polymer for Cryopreservation

Sun, Yuling; Liu, Jie; Li, Zhibo; Wang, Jianjun; Huang, Yanbin

doi:10.1021/acsami.0c22308

Cited by 20 publications

(22 citation statements)

References 43 publications

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“…Therefore, cryopreservation cannot simply be attributed to ice recrystallization inhibition, as opposed to other effects (especially when high concentrations are used). To highlight this, poly( D / L -serine) and poly(ethylene glycol) (PEG), which have limited ice recrystallization inhibitory activity, can aid red blood cell cryopreservation at high concentrations (100 mg ml −1 ) 55 . By contrast, oligoproline, which is a weak IRI, is a potent cryopreservation enhancer for monolayers of A549 cells 56 (at ~10 mg ml −1 ) and oocytes 57 .…”

Section: Chemical Tools For Cryopreservationmentioning

confidence: 99%

Chemical approaches to cryopreservation

2022

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Cryopreservation of cells and biologics underpins all biomedical research from routine sample storage to emerging cell-based therapies, as well as ensuring cell banks provide authenticated, stable and consistent cell products. This field began with the discovery and wide adoption of glycerol and dimethyl sulfoxide as cryoprotectants over 60 years ago, but these tools do not work for all cells and are not ideal for all workflows. In this Review, we highlight and critically review the approaches to discover, and apply, new chemical tools for cryopreservation. We summarize the key (and complex) damage pathways during cellular cryopreservation and how each can be addressed. Bio-inspired approaches, such as those based on extremophiles, are also discussed. We describe both small-molecule-based and macromolecular-based strategies, including ice binders, ice nucleators, ice nucleation inhibitors and emerging materials whose exact mechanism has yet to be understood. Finally, looking towards the future of the field, the application of bottom-up molecular modelling, library-based discovery approaches and materials science tools, which are set to transform cryopreservation strategies, are also included.

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Section: Chemical Tools For Cryopreservationmentioning

confidence: 99%

Chemical approaches to cryopreservation

2022

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“…When the PEG-PAK T concentration was fixed at 0.10 wt % and the trehalose concentration varied over 0.00–15.0 wt %, the ice crystal size significantly decreased (Figure c). The IRI activity was quantitatively assayed by mean largest grain size (MLGS), , which indicated that ice crystal size was controlled dominantly by the trehalose concentration. When the PEG-PAK T concentration was fixed at 0.10 wt % and the trehalose concentration increased from 0.00 to 15.0 wt %, the MLGS decreased from 76 to 28% of that of PBS.…”

Section: Resultsmentioning

confidence: 99%

“…Cryomicroscopy images of the wafer were collected through crossed polarizers using a microscope (CX40IT; Soptop, China) equipped with a 10×/0.25/∞/-/FN26.5 lens (UIS-2; Olympus Ltd, Japan) and a 3 M CMOS Color Digital Camera (BoliOptics). The mean largest grain size (MLGS) of ice crystals, that is the size of the 10 largest crystals from each wafer, was measured using ImageJ and expressed as a relative size to MLGS of ice crystals formed from PBS …”

Section: Methodsmentioning

confidence: 99%

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Poly(l-alanine-co-l-lysine)-g-Trehalose as a Biomimetic Cryoprotectant for Stem Cells

et al. 2022

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Poly(L-alanine-co-L-lysine)-graft-trehalose (PAK T ) was synthesized as a natural antifreezing glycopolypeptide (AFGP)-mimicking cryoprotectant for cryopreservation of mesenchymal stem cells (MSCs). FTIR and circular dichroism spectra indicated that the content of the α-helical structure of PAK decreased after conjugation with trehalose. Two protocols were investigated in cryopreservation of MSCs to prove the significance of the intracellularly delivered PAK T . In protocol I, MSCs were cryopreserved at −196 °C for 7 days by a slow-cooling procedure in the presence of both PAK T and free trehalose. In protocol II, MSCs were preincubated at 37 °C in a PAK T solution, followed by cryopreservation at −196 °C in the presence of free trehalose for 7 days by the slow-cooling procedure. Polymer and trehalose concentrations were varied by 0.0−1.0 and 0.0−15.0 wt %, respectively. Cell recovery was significantly improved by protocol II with preincubation of the cells in the PAK T solution. The recovered cells from protocol II exhibited excellent proliferation and maintained multilineage potentials into osteogenic, chondrogenic, and adipogenic differentiation, similar to MSCs recovered from cryopreservation in the traditional 10% dimethyl sulfoxide system. Ice recrystallization inhibition (IRI) activity of the polymers/trehalose contributed to cell recovery; however, intracellularly delivered PEG-PAK T was the major contributor to the enhanced cell recovery in protocol II. Inhibitor studies suggested that macropinocytosis and caveolin-dependent endocytosis are the main mechanisms for the intracellular delivery of PEG-PAK T . 1 H NMR and FTIR spectra suggested that the intracellular PEG-PAK T s interact with water and stabilize the cells during cryopreservation.

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“…20−22 As an analogue of the hydroxyl group of PVA, hydroxy nylon-3 and DL-polyserine were reported as cryoprotecting compounds. 23,24 However, their IRI activities were much lower than that of PVA. On the other hand, atactic PVA exhibited better IRI activity than isotactic PVA, whose aqueous solubility limits their IRI activity.…”

Section: ■ Introductionmentioning

confidence: 94%

Ice Recrystallization Inhibition Using l-Alanine/l-Lysine Copolymers

Park

Piao

Park

et al. 2022

ACS Appl. Polym. Mater.

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Recently, poly(L-alanine-co-L-lysine) (PAK) was reported to exhibit an ice recrystallization inhibition (IRI) activity comparable to that of poly(vinyl alcohol), which has the highest IRI activity among synthetic polymers (ACS Macro Lett. 2021, 10, 1436−1442. As a continuation of the study, we synthesized a series of PAK copolymers to investigate the structure−property relationship of PAK on the IRI activity and its mechanism by varying the (1) composition of the L-alanine/L-lysine (A/K) ratio among 0/100, 50/50, 60/40, and 70/30 and (2) molecular weight of PAK among 4, 9, and 15 kDa at a fixed 50/50 ratio of A/K. As the hydrophobic L-alanine composition of PAK increased from 0 to 70%, the mean largest grain size (MLGS) relative to phosphatebuffered saline decreased from 74 to 11% at 1.0 mg/mL. As the molecular weight of PAK increased from 4 to 15 kDa, the MLGS decreased from 32 to 15% at 1.0 mg/mL. The ice nucleation temperature decreased as the hydrophobic L-alanine composition and the molecular weight of PAK increased. The thermal hysteresis (TH) of PAK was about 0.05 °C at 1.0 mg/mL and increased to 0.2 °C as the molecular weight increased from 4 to 15 kDa. The ice crystal size of ice cream showed a similar trend to IRI activity. To understand the underlying chemistry of the IRI effect of PAK, circular dichroism spectroscopy, a dynamic ice shaping study, and Fourier-transform infrared spectroscopy were carried out. These studies suggest that the α-helicity, hydrophobic/hydrophilic balance, molecular weight of PAK, and extent of interactions between polymers and ice crystals are related to the IRI activity, which extends to ice nucleation inhibition, TH of ice formation, ice shaping, and even the ice crystal size of ice cream.

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Nonionic and Water-Soluble Poly(d/l-serine) as a Promising Biomedical Polymer for Cryopreservation

Cited by 20 publications

References 43 publications

Chemical approaches to cryopreservation

Chemical approaches to cryopreservation

Poly(l-alanine-co-l-lysine)-g-Trehalose as a Biomimetic Cryoprotectant for Stem Cells

Ice Recrystallization Inhibition Using l-Alanine/l-Lysine Copolymers

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