Characterization of injury in isolated rat proximal tubules during cold incubation and rewarming

Bienholz, Anja; Walter, Britta; Pless-Petig, Gesine; Guberina, Hana; Kribben, Andreas; Witzke, Oliver; Rauen, Ursula

doi:10.1371/journal.pone.0180553

Cited by 12 publications

(15 citation statements)

References 45 publications

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“…During cold storage, function of ion pumps like the Na + /K + -ATPase is reduced, which has been suggested to impede maintenance of ion equilibrium leading to cell swelling and necrosis (42). However, in the presence of iron chelators, cold storage in chloride-rich preservation solutions, with chloride concentrations comparable to cell culture medium, proved to be superior to cold storage in chloride-poor organ preservation solutions for cell types such as rat proximal tubules and porcine aortic endothelial cells (26,43).…”

Section: Discussionmentioning

confidence: 99%

Optimization of long-term cold storage of rat precision-cut lung slices with a tissue preservation solution

Tigges¹,

Eggerbauer²,

Worek³

et al. 2021

American Journal of Physiology-Lung Cellular and Molecular Phys

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Precision cut lung slices (PCLS) are used as ex vivo model of the lung to fill the gap between in vitro and in vivo experiments. To allow optimal utilization of PCLS, possibilities to prolong slice viability via cold storage using optimized storage solutions were evaluated. Rat PCLS were cold stored in DMEM/F-12 or two different preservation solutions for up to 28 days at 4 °C. After rewarming in DMEM/F-12, metabolic activity, live/dead staining and mitochondrial membrane potential was assessed to analyze overall tissue viability. Single cell suspensions were prepared and proportions of CD45+, EpCAM+, CD31+ and CD90+ cells analyzed. As functional parameters, TNF-α expression was analyzed to detect inflammatory activity and bronchoconstriction after acetylcholine stimulus. After 14 days cold storage, viability and mitochondrial membrane potential were significantly better preserved after storage in solution 1 (potassium chloride rich) and solution 2 (potassium- and lactobionate-rich analog) compared to DMEM/F-12. Analysis of cell populations revealed efficient preservation of EpCAM+, CD31+ and CD90+ cells. Proportion of CD45+ cells decreased during cold storage but was better preserved by both modified solutions than by DMEM/F-12. PCLS stored in solution 1 responded substantially longer to inflammatory stimulation than those stored in DMEM/F-12 or solution 2. Analysis of bronchoconstriction revealed total loss of function after 14 days storage in DMEM/F-12 but in contrast, a good response in PCLS stored in the optimized solutions. An improved base solution with a high potassium chloride concentration optimizes cold storage of PCLS and allows shipment between laboratories and stockpiling of tissue samples.

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Section: Discussionmentioning

confidence: 99%

Optimization of long-term cold storage of rat precision-cut lung slices with a tissue preservation solution

Tigges¹,

Eggerbauer²,

Worek³

et al. 2021

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…10 The high potassium concentration protected against the loss of the mitochondrial membrane potential. 10 For the cold storage of rat renal proximal tubules, extracellular chloride has recently been shown to protect mitochondria, 51 although the exact mechanism of mitochondrial protection is still unclear. In contrast to porcine aortic endothelial cells and rat renal proximal tubules, rat hepatocytes display a chloride-dependent cell injury after cold (4°C) storage in chloride-rich solutions and are therefore significantly better protected in chloride-poor solutions.…”

Section: Discussionmentioning

confidence: 99%

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

2018

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Cryopreservation is the only long-term storage option for the storage of vessels and vascular constructs. However, endothelial barrier function is almost completely lost after cryopreservation in most established cryopreservation solutions. We here aimed to improve endothelial function after cryopreservation using the 2D-model of porcine aortic endothelial cell monolayers. The monolayers were cryopreserved in cell culture medium or cold storage solutions based on the 4°C vascular preservation solution TiProtec®, all supplemented with 10% DMSO, using different temperature gradients. After short-term storage at −80°C, monolayers were rapidly thawed and re-cultured in cell culture medium. Thawing after cryopreservation in cell culture medium caused both immediate and delayed cell death, resulting in 11 ± 5% living cells after 24 h of re-culture. After cryopreservation in TiProtec and chloride-poor modifications thereof, the proportion of adherent viable cells was markedly increased compared to cryopreservation in cell culture medium (TiProtec: 38 ± 11%, modified TiProtec solutions ≥ 50%). Using these solutions, cells cryopreserved in a sub-confluent state were able to proliferate during re-culture. Mitochondrial fragmentation was observed in all solutions, but was partially reversible after cryopreservation in TiProtec and almost completely reversible in modified solutions within 3 h of re-culture. The superior protection of TiProtec and its modifications was apparent at all temperature gradients; however, best results were achieved with a cooling rate of −1°C/min. In conclusion, the use of TiProtec or modifications thereof as base solution for cryopreservation greatly improved cryopreservation results for endothelial monolayers in terms of survival and of monolayer and mitochondrial integrity.

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“…For isolated hepatocytes, storage times of 7 d are already possible . However, mitochondrial function/ATP generation might be compromised in cells after prolonged cold storage . Further analysis of these limitations is likely to identify additional targets for intervention, thus allowing to further improve cold preservation and extend cold preservation periods.…”

Section: Discussionmentioning

confidence: 99%

Preservation of Cell Structure, Metabolism, and Biotransformation Activity of Liver‐On‐Chip Organ Models by Hypothermic Storage

Gröger

Dinger

Kiehntopf

et al. 2017

Adv Healthcare Materials

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The liver is a central organ in the metabolization of nutrition, endogenous and exogenous substances, and xenobiotic drugs. The emerging organ-on-chip technology has paved the way to model essential liver functions as well as certain aspects of liver disease in vitro in liver-on-chip models. However, a broader use of this technology in biomedical research is limited by a lack of protocols that enable the short-term preservation of preassembled liver-on-chip models for stocking or delivery to researchers outside the bioengineering community. For the first time, this study tested the ability of hypothermic storage of liver-on-chip models to preserve cell viability, tissue morphology, metabolism and biotransformation activity. In a systematic study with different preservation solutions, liver-on-chip function can be preserved for up to 2 d using a derivative of the tissue preservation solution TiProtec, containing high chloride ion concentrations and the iron chelators LK614 and deferoxamine, supplemented with polyethylene glycol (PEG). Hypothermic storage in this solution represents a promising method to preserve liver-on-chip function for at least 2 d and allows an easier access to liver-on-chip technology and its versatile and flexible use in biomedical research.

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Characterization of injury in isolated rat proximal tubules during cold incubation and rewarming

Cited by 12 publications

References 45 publications

Optimization of long-term cold storage of rat precision-cut lung slices with a tissue preservation solution

Optimization of long-term cold storage of rat precision-cut lung slices with a tissue preservation solution

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

Preservation of Cell Structure, Metabolism, and Biotransformation Activity of Liver‐On‐Chip Organ Models by Hypothermic Storage

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