Influence of intracellular trehalose concentration and pre-freeze cell volume on the cryosurvival of rapidly frozen human erythrocytes

Lynch, Andrew; Slater, Nigel K.H.

doi:10.1016/j.cryobiol.2011.04.005

Cited by 25 publications

(25 citation statements)

References 23 publications

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“…The latter two methods also have the disadvantage of requiring incubation times of 24 h or more. This is in contrast to the proposed method of PP-50 mediated trehalose delivery [27].…”

Section: Introductionmentioning

confidence: 64%

“…Recently, the amphipathic membrane permeabilising polymer PP-50 has been used to load human erythrocytes with trehalose, which led to a significant enhancement in cryosurvival [27]. PP-50, which can be removed from cell membranes by a small change in pH [26], is thought to be non-cytotoxic [11,22].…”

Section: Introductionmentioning

confidence: 99%

“…In the current study, the techniques for the cryopreservation of cells using trehalose and PP-50 developed by Lynch et al [27] were extended to successfully preserve nucleated human cells. The Human osteosarcoma derived cell line SAOS-2 [16,35] was used as a model for nucleated, adherent human cells.…”

Section: Introductionmentioning

confidence: 99%

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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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“…The latter two methods also have the disadvantage of requiring incubation times of 24 h or more. This is in contrast to the proposed method of PP-50 mediated trehalose delivery [27].…”

Section: Introductionmentioning

confidence: 64%

Section: Introductionmentioning

confidence: 99%

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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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“…[10][11][12] Suggested method of trehalose internalization was by fluid phase endocytosis after 1-24 hours of trehalose pre-treatment via clathrin-mediated endocytosis. 12 However, this was suggested as cell-type specific because the endocytosis mechanism involved.…”

Section: Discussionmentioning

confidence: 99%

Trehalose preincubation increases mesenchymal (CD271<sup>+</sup>) stem cells post-cryopreservation viability

et al. 2016

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ABSTRAK ABSTRACTBackground: Dimethyl sulfoxide (Me2SO) is a common cryoprotective agent widely used in cell preservation system. Me2SO is currently known to cause epigenetic changes which are critical in stem cells development and cellular differentiation. Therefore, it is imperative to develop cryopreservation techniques that protect cellular functions and avert Me2SO adverse effect. Trehalose was able to protect organism in extreme condition such as dehydration and cold. This study aimed to verify the protective effect of trehalose preincubation procedure in cryopreservation.

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Influence of intracellular trehalose concentration and pre-freeze cell volume on the cryosurvival of rapidly frozen human erythrocytes

Cited by 25 publications

References 23 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

Trehalose preincubation increases mesenchymal (CD271<sup>+</sup>) stem cells post-cryopreservation viability

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