On crystallization of water confined in liposomes and cryoprotective action of DMSO

Klbik, Ivan; Čechová, Katarína; Maťko, I.; Lakota, Ján; Šauša, O.

doi:10.1039/d1ra08935h

Cited by 10 publications

(7 citation statements)

References 53 publications

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“…Given the broadening of the endothermic peak and the changes in T m and Δ H fus as shown in Figure d, e, we speculate that the unfrozen water content of the OE 2 imC 3 C solution is higher than that of the two CPA solutions. The fraction of unfrozen water ( U ) in each solution was determined using the following equation U = 1 − normalΔ H exp normalΔ H 0 Here, Δ H exp represents the experimentally determined enthalpic change during ice melting, normalized to the weight of water in the solution.…”

Section: Resultsmentioning

confidence: 99%

Protein Cryoprotectant Ability of the Aqueous Zwitterionic Solution

Takekiyo,

Yamada,

Uto

et al. 2024

J. Phys. Chem. B

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Protein cryopreservation is important for the long-term storage of unstable proteins. Recently, we found that N-acetylglucosaminyltransferase-V (GnT-V) can be cryopreserved in a deep freezer without temperature control using a dilute binary aqueous solution of 3-(1-(2-(2-methoxyethoxy)ethyl)imidazol-3-io)butane-1-carboxylate (OE 2 imC 3 C) [10 wt %, mole fraction of solute (x) = 7.75 × 10 −3 ], an artificial zwitterion. However, it is unclear which solvent properties are required in these media to preserve unstable proteins, such as GnT-V. In this study, we investigated the melting phenomena and solution structure of dilute binary aqueous OE 2 imC 3 C solutions [x = 0−2.96 × 10 −2 (0−30 wt %)] using differential scanning calorimetry (DSC) and Raman and Fourier transform infrared (FTIR) spectroscopies combined with molecular dynamics (MD) simulation to compare the cryoprotectant ability of OE 2 imC 3 C with two general cryoprotectants (CPAs), glycerol and dimethyl sulfoxide. DSC results indicated that aqueous OE 2 imC 3 C solutions can be melted at lower temperatures with less energy than the control CPA solution, with increasing x, primarily due to OE 2 imC 3 C having a higher content of unfrozen water molecules. Moreover, Raman and FTIR results showed that the high content of unfrozen water molecules in aqueous OE 2 imC 3 C solutions was due to the hydration around the ionic parts (the COO − group and imidazolium ring) and the OCH 2 CH 2 O segment. In addition, the MD simulation results showed that there were fewer structured water molecules around the OCH 2 CH 2 O segment than the hydration water molecules around the ionic parts. These solvent properties suggest that dilute aqueous OE 2 imC 3 C solutions are effective in preventing freezing, even in a deep freezer. Therefore, this medium has the potential to act as a novel cryoprotectant for proteins in biotechnology and biomedical fields.

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Section: Resultsmentioning

confidence: 99%

Protein Cryoprotectant Ability of the Aqueous Zwitterionic Solution

Takekiyo,

Yamada,

Uto

et al. 2024

J. Phys. Chem. B

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“…Interestingly, notwithstanding Me 2 SO can contribute to the amorphous phase formation, DSC and, especially, TMA, Me 2 SO alone cannot inhibit the process. The amorphous phase formation is generally considered a good prognostic factor for promoting cell survi val [35]. However, the work has shown that not all polymers were equally effective in terms of cell recovery after cryopreservation.…”

Section: Resultsmentioning

confidence: 99%

Study of physical processes occurring in serum-containing and polymer-based serum-free cryoprotective media

Pakhomov,

Gurina,

Polyakova

et al. 2024

Biopolym. Cell

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To investigate the mechanisms of cryoprotection with compositions that were supplemented with polymers (dextran [Dex], hydroxyethyl starch [HES], polyethylene glycol [PEG]) or fetal bovine serum [FBS]). Methods. Differential scanning calorimetry (DSC) and thermomechanical analysis (TMA) of cryoprotective media containing serum, as well as media that do not contain serum or its components but include polymers. Results. The combinations of dimethyl sulfoxide with FBS or polymers in cryoprotective media were able to promote the formation of amorphous phase and suppress salt eutectic crystallization/mel ting and recrystallization.The medium supplemented with 0.7 M Me 2 SO and 100 mg/ml Dex (0.7Me 2 SO+Dex) was somewhat superior compared to other compositions. It also had an optimal glass transition temperature (-26…-21 °C). The feature correlated with an adequate cell recovery when the medium was utilized to preserve testicular cells. Conclusions. The mechanism of cryoprotection depends not only on the concentrations of cryoprotective agents such as Me 2 SO, polymers or FBS, but also on the composition of the cryoprotective media. The 0.7Me 2 SO+Dex medium can be the basis for the creation of commercial serum-free cryoprotective media with a defined stable composition that does not require removal of Me 2 SO before using the cryopreserved cells or tissues for cultivation, transplantation and transfusion.

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“…Similar depressions of nucleation and melting temperature were reported in some works for water-Me 2 SO systems. 37 The effect of 10% FBS or 100 mg/mL of poly-mers alone (without Me 2 SO) on the melting temperature of Ham's F12 was undetectable by cryomicroscopy itself (Table 3).…”

Section: Ta B L Ementioning

confidence: 97%

Open‐source hardware‐ and software‐based cryomicroscopy system for investigation of phase transitions in cryobiological research

Pakhomov,

Shevchenko,

Chernobai

et al. 2023

Journal of Microscopy

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The development of inexpensive equipment adapted for the study of a specific biological object is very important for cryobiology. In the presented work, we have proposed a simple system for microscopy utilising open‐source platform Arduino. Testing this system showed that it had sufficient sensitivity to determine the physical processes occurring in a cryopreserved sample such as intra‐ and extracellular water crystallisation and salt eutectic. Utilising this system, we investigated the mechanisms of cryoprotection and cryodamage of testis interstitial cells (ICs) in cryoprotective media, which included cryoprotective agents such as dimethyl sulphoxide (Me2SO), as well as foetal bovine serum or polymers (dextran, hydroxyethyl starch and polyethylene glycol). It was shown that a serum‐/xeno‐free medium that included 0.7 M Me2SO and 100 mg/mL dextran was able to reduce intracellular water crystallisation in cells, change the structure of extracellular ice, and reduce salt eutectic and recrystallisation. All these effects correlated with better IC survival after cryopreservation in the medium. This medium is potentially less toxic as it has lower concentrations of Me2SO compared to serum‐containing media developed for cryopreservation of testicular cells. This would pave a way for the creation of nontoxic serum‐free compositions that does not require removal before use of cryopreserved living cells for laboratory practice or in clinics.

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On crystallization of water confined in liposomes and cryoprotective action of DMSO

Abstract: Ice-free phase formed by DMSO acting as a protective layer of lipid membrane.

Cited by 10 publications

References 53 publications

Protein Cryoprotectant Ability of the Aqueous Zwitterionic Solution

Protein Cryoprotectant Ability of the Aqueous Zwitterionic Solution

Study of physical processes occurring in serum-containing and polymer-based serum-free cryoprotective media

Open‐source hardware‐ and software‐based cryomicroscopy system for investigation of phase transitions in cryobiological research

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