Protective effects of osmolytes in cryopreserving adherent neuroblastoma (Neuro-2a) cells

Bailey, Trisha L.; Wang, Mian; Solocinski, Jason; Nathan, Britto P.; Chakraborty, Nilay; Menze, Michael A.

doi:10.1016/j.cryobiol.2015.08.015

Cited by 36 publications

(43 citation statements)

References 51 publications

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“…This outcome is consistent with previous studies, which found that sugars suppressed the growth rate of ice crystals due to the strong hydrogen bonding of sugar with water molecules (Sei, Gonda, & Arima, 2002;Uchida, Nagayama, Shibayama, & Gohara, 2007). These results are also consistent with the findings of Bailey et al (2015), who found that the addition of trehalose to DMSO changed the ice crystal patterns observed upon freezing.…”

Section: Sugarssupporting

confidence: 93%

“…Sugar alcohols can also interact with water, and alteration of ice structure by osmolytes has been demonstrated to be influential in cryopreservation outcome (Bailey et al, ). To examine the interactions between osmolytes and water during freezing, Raman images of osmolyte solutions without cells cooled down to −50°C with cooling rate 1°C/min were obtained for SGI353 (sucrose–glycerol–isoleucine; Pi et al, ), SGC353, TGC353, and SMC353 solutions.…”

Section: Resultsmentioning

confidence: 99%

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Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells

Dosa

et al. 2019

Biotech & Bioengineering

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This study examined the post‐thaw recovery of Jurkat cells cryopreserved in three combinations of five osmolytes including trehalose, sucrose, glycerol, mannitol, and creatine. Cellular response was characterized using low‐temperature Raman spectroscopy, and variation of post‐thaw recovery was analyzed using statistical modeling. Combinations of osmolytes displayed distinct trends of post‐thaw recovery, and a nonlinear relationship between compositions and post‐thaw recovery was observed, suggesting interactions not only between different solutes but also between solutes and cells. The post‐thaw recovery for optimized cryoprotectants in different combinations of osmolytes at a cooling rate of 1°C/min was comparable to that measured with 10% dimethyl sulfoxide. Statistical modeling was used to understand the importance of individual osmolytes as well as interactions between osmolytes on post‐thaw recovery. Both higher concentrations of glycerol and certain interactions between sugars and glycerol were found to typically increase the post‐thaw recovery. Raman images showed the influence of osmolytes and combinations of osmolytes on ice crystal shape, which reflected the interactions between osmolytes and water. Differences in the composition also influenced the presence or absence of intracellular ice formation, which could also be detected by Raman. These studies help us understand the modes of action for cryoprotective agents in these osmolyte solutions.

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Section: Sugarssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells

Dosa

et al. 2019

Biotech & Bioengineering

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“…Studies suggest that sugars interact directly with the cell membrane during freezing [34, 35]. The addition of sugars to solutions containing DMSO also results in changes in ice patterns that are observed [36]. Sugars such as trehalose and sucrose have gained popularity in the cryopreservation of HSCs and satisfying cryopreservation outcomes have been broadly demonstrated.…”

Section: New Cryoprotectantsmentioning

confidence: 99%

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

Hornberger

McKenna

et al. 2019

Transfus Med Hemother

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Hematopoietic stem cell (HSC) therapy is widely used to treat a growing number of hematological and non-hematological diseases. Cryopreservation of HSCs allows for cells to be transported from the site of processing to the site of clinical use, creates a larger window of time in which cells can be administered to patients, and allows sufficient time for quality control and regulatory testing. Currently, HSCs and other cell therapies conform to the same cryopreservation techniques as cells used for research purposes: cells are cryopreserved in dimethyl sulfoxide (DMSO) at a slow cooling rate. As a result, HSC therapy can result in numerous adverse symptoms in patients due to the infusion of DMSO. Efforts are being made to improve the cryopreservation of HSCs for clinical use. This review discusses advances in the cryopreservation of HSCs from 2007 to the present. The preclinical development of new cryoprotectants and new technology to eliminate cryoprotectants after thawing are discussed in detail. Additional cryopreservation considerations are included, such as cooling rate, storage temperature, and cell concentration. Preclinical cell assessment and quality control are discussed, as well as clinical studies from the past decade that focus on new cryopreservation protocols to improve patient outcomes.

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“…Growth of cells in medium supplemented with trehalose allowing trehalose uptake via fluid‐phase endocytosis prior to freezing has been shown to increase cryosurvival . The mechanisms have been attributed to metabolic downregulation by trehalose or osmotic stress caused by culturing with trehalose activating signaling pathways that allow cells to better cope with cryopreservation stress.…”

Section: Discussionmentioning

confidence: 99%

“…30 Fluid-phase endocytosis, which is an inherent property of cells to incorporate membrane-impermeable molecules, can also be used to introduce trehalose into cells. 31 It has been shown that growth of cells in the presence of trehalose, allowing uptake through endocytosis, increases cell stability during cryopreservation [32][33][34] and freeze-drying. 31 Trehalose uptake via fluid-phase endocytosis shows a linear dependency with respect to the extracellular concentration and incubation time and is inhibited at low temperatures.…”

Section: Introductionmentioning

confidence: 99%

Combining endocytic and freezing‐induced trehalose uptake for cryopreservation of mammalian cells

Zhang

Oldenhof

Sieme

et al. 2016

Biotechnology Progress

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Fibroblasts take up trehalose during freezing and thawing, which facilitates cryosurvival of the cells. The aim of this study was to investigate if trehalose uptake via fluid-phase endocytosis prefreeze increases cryosurvival. To determine endocytic trehalose uptake in attached as well as suspended fibroblasts, intracellular trehalose concentrations were determined during incubation at 37°C using an enzymatically based trehalose assay. In addition, freezing-induced trehalose uptake of extracellularly added trehalose was determined. Cryosurvival rates were determined via trypan blue staining. Intracellular trehalose contents of attached as well as suspended cells were found to increase linearly with time, consistent with fluid-phase endocytosis. Furthermore, the intracellular trehalose concentration increased with increasing extracellular trehalose concentration (0-100 mM) in a linear fashion. Prefreeze loading of cells with trehalose via fluid-phase endocytosis only showed increased cryosurvival rates at extracellular trehalose concentrations lower than 50 mM in the cryopreservation medium. To obtain satisfactory cryosurvival rates after endocytic preloading, extracellular trehalose is needed to prevent efflux of trehalose during freezing and thawing and for freezing-induced trehalose uptake. At trehalose concentrations greater than 100 mM, cryosurvival rates were similar or slightly higher if cells were not loaded with trehalose prefreeze. Cells that were grown in the presence of trehalose showed a tendency to aggregate after harvesting. It is concluded that it is particularly freezing-induced trehalose uptake that facilitates cryosurvival when trehalose is used as the sole cryoprotectant for cryopreservation of fibroblasts. Preloading with trehalose does not increase cryosurvival rates if trehalose is also added as extracellular protectant. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:229-230, 2017.

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Protective effects of osmolytes in cryopreserving adherent neuroblastoma (Neuro-2a) cells

Cited by 36 publications

References 51 publications

Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells

Characterizing modes of action and interaction for multicomponent osmolyte solutions on Jurkat cells

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

Combining endocytic and freezing‐induced trehalose uptake for cryopreservation of mammalian cells

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