Cryopreservation of hepatocyte (HepG2) cell monolayers: Impact of trehalose

“…Trehalose, a macromolecular substance, greatly affects cells osmotic pressure. Trehalose on cryopreservation could protect the cell and tissue activity (Lee et al, ; Stokich et al, ; Zhang, Hu, et al, ). Previous studies suggest that trehalose, in a dose‐dependent manner has different effects on cryopreservation of bovine semen (Hu et al, ), boar testes (Lee et al, ) and bovine calf testes (Zhang, Wang, et al, ).…”

Section: Discussionmentioning

confidence: 99%

“…Trehalose improves mammalian cells’ ability to withstand cold temperature (Eroglu et al, ). Incubation with trehalose is beneficial for long‐term preservation of cells (Stokich et al, ). Trehalose is also applied as an antioxidant in frozen–thawed bovine calf testicular tissue (Zhang, Hu, et al, ).…”

Section: Introductionmentioning

confidence: 99%

Trehalose protects testicular tissue of dairy goat upon cryopreservation

Hao

Ren

Zhang

et al. 2019

Reprod Domestic Animals

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The aim of this study was to investigate whether the addition of trehalose to cryomedia reduces cellular damage and improves gene expression in cryopreserved dairy goat testicular tissues. Testicular tissues were cryopreserved in cryomedia without or with trehalose at a concentration of 5%, 10%, 15%, 20% or 25%. Cryopreserved testicular tissues were analysed for TUNEL‐positive cell number, expression of BAX, BCL‐2, CREM, BOULE and HSP70‐2. Isolated Leydig cells from cryopreserved tissue were cultured, and spent medium was evaluated for testosterone level. The results showed that though the TUNEL‐positive cell number increased in cryopreserved testicular tissues, the presence of trehalose reduced apoptotic cell number significantly. Quantitative real‐time polymerase chain reaction results showed that although the expression of BAX was upregulated following cryopreservation, the presence of trehalose downregulates it in cryopreserved testicular tissues. Expression of BCL‐2, CREM, BOULE and HSP70‐2 was downregulated following cryopreservation but the presence of trehalose significantly upregulated their expression in cryopreserved testicular tissues. Leydig cells isolated from testicular tissues cryopreserved with trehalose produced higher testosterone than the one without it (control). These results suggest that trehalose has a protective role in cryopreservation of dairy goat testicular tissue, and the most suitable trehalose concentration for cryopreservation is 15%.

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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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Cryopreservation of hepatocyte (HepG2) cell monolayers: Impact of trehalose

Abstract: 23A simple method to cryogenically preserve hepatocyte monolayers is currently not available but

Cited by 47 publications

References 57 publications

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

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

Trehalose protects testicular tissue of dairy goat upon cryopreservation

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

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