Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival

Lynch, Andrew; Chen, Rongjun; Dominowski, Paul J.; Shalaev, Evgenyi; Yancey, Robert J.; Slater, Nigel K.H.

doi:10.1016/j.biomaterials.2010.04.020

Cited by 71 publications

(113 citation statements)

References 37 publications

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“…Materials for the PP-50 polymer synthesis were sourced as previously described [25]. Foetal bovine serum (FBS), l -glutamine, and penicillin/streptomycin were purchased from Invitrogen (UK).…”

Section: Methodsmentioning

confidence: 99%

Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation

et al. 2013

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For stem cell therapy to become a routine reality, one of the major challenges to overcome is their storage and transportation. Currently this is achieved by cryopreserving cells utilising the cryoprotectant dimethyl sulfoxide (Me2SO). Me2SO is toxic to cells, leads to loss of cell functionality, and can produce severe side effects in patients. Potentially, cells could be frozen using the cryoprotectant trehalose if it could be delivered into the cells at a sufficient concentration. The novel amphipathic membrane permeabilising agent PP-50 has previously been shown to enhance trehalose uptake by erythrocytes, resulting in increased cryosurvival. Here, this work was extended to the nucleated human cell line SAOS-2. Using the optimum PP-50 concentration and media osmolarity, cell viability post-thaw was 60 ± 2%. In addition, the number of metabolically active cells 24 h post-thaw, normalised to that before freezing, was found to be between 103 ± 4% and 91 ± 5%. This was found to be comparable to cells frozen using Me2SO. Although reduced (by 22 ± 2%, p = 0.09), the doubling time was found not to be statistically different to the non-frozen control. This was in contrast to cells frozen using Me2SO, where the doubling time was significantly reduced (by 41 ± 4%, p = 0.004). PP-50 mediated trehalose delivery into cells could represent an alternative cryopreservation protocol, suitable for research and therapeutic applications.

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Section: Methodsmentioning

confidence: 99%

Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation

et al. 2013

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“…However, removing the high concentration of glycerol before transfusion increased operative time and costs. Th us, several scholars focused their eff orts on this issue and reported that the high concentration of glycerol could be replaced by the oligosaccharide (Quan et al 2011), trehalose (Lynch and Slater 2011), or biopolymers (Lynch et al 2010), which can be easily removed. However, the addition of such a high concentration of cryoprotectant during the erythrocytes cryopreservation results in high osmolality (5000 mOsm/kg) that is almost 17-fold higher than the normal physiological osmolality (Robert Valeri and Ragno 2007).…”

Section: Introductionmentioning

confidence: 99%

Influence of a static magnetic field on the slow freezing of human erythrocytes

Lin

Chang

Lee

et al. 2012

International Journal of Radiation Biology

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“…However, trehalose alone cannot work well in cryopreservation due to its poor permeability into the cells. Several reports have shown that its cryoprotect efficacy is improved if it can be delivered into the cells with various strategies, such as microinjection, pore formation using mutant bacterial toxins, or internal trehalose synthesis via genic engineering, etc 333435…”

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confidence: 99%

Natural zwitterionic betaine enables cells to survive ultrarapid cryopreservation

Yang

Cai

Zhai

et al. 2016

Sci Rep

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Cryoprotectants (CPAs) play a critical role in cryopreservation because they can resist the cell damage caused by the freezing process. Current state-of-the-art CPAs are mainly based on an organic solvent dimethyl sulfoxide (DMSO), and several DMSO-cryopreserved cell products have been brought to market. However, the intrinsic toxicity and complex freezing protocol of DMSO still remain as the bottleneck of the wide use for clinical applications. Herein, we reported that betaine, a natural zwitterionic molecule, could serve as a nontoxic and high efficient CPA. At optimum concentration of betaine, different cell types exhibited exceptional post-thaw survival efficiency with ultrarapid freezing protocol, which was straightforward, cost efficient but difficult to succeed using DMSO. Moreover, betaine showed negligible cytotoxicity even after long-term exposure of cells. Mechanistically, we hypothesized that betaine could be ultra-rapidly taken up by cells for intracellular protection during the freezing process. This technology unlocks the possibility of alternating the traditional toxic CPAs and is applicable to a variety of clinical applications.

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Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival

Cited by 71 publications

References 37 publications

Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation

Amphipathic polymer-mediated uptake of trehalose for dimethyl sulfoxide-free human cell cryopreservation

Influence of a static magnetic field on the slow freezing of human erythrocytes

Natural zwitterionic betaine enables cells to survive ultrarapid cryopreservation

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