Freezing Properties of Disaccharide Solutions: Inhibition of Hexagonal Ice Crystal Growth and Formation of Cubic Ice

Uchida, Tsutomu; Takeya, Satoshi; Nagayama, Masaichi; Gohara, Kazutoshi

doi:10.5772/29694

Cited by 8 publications

(7 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This trend indicates that the CPA effect by extracellular trehalose is expected to be higher at higher trehalose concentrations. Such an effect appears consistent with several proposed cryoprotective mechanisms from extracellular trehalose: both of which inhibits intracellular freezing [15,[44][45][46]. And (iii), trehalose interacts directly with cell membranes and proteins and protecting them [5-6, 25, 31, 40, 42-43].…”

Section: Freeze-thaw Viabilities Of Cho-vector Cells and Their Apoptosupporting

confidence: 82%

Trehalose uptake and dehydration effects on the cryoprotection of CHO–K1 cells expressing TRET1

et al. 2019

Self Cite

View full text Add to dashboard Cite

Chinese hamster ovary cells (CHO-K1 cells) in which the trehalose transporter (TRET1) is expressed can have greater cryoprotection than ordinary CHO-K1 cells. This study examines the uptake characteristics of trehalose into cells via TRET1 and determines the influence of intracellular trehalose on the freeze-thaw viabilities. In our experiments, the intracellular trehalose concentration is controlled by the extracellular trehalose concentration and the immersion time in a freezing solution. In this freezing solution, both kinds of CHO-K1 cells are independently dispersed with various amount of trehalose, and then put into the CO2 incubator for 0 to 6 h. After a set immersion time, the cell-suspended sample is cooled to 193 K, stored for 1 week, then quickly thawed at 310 K and its viability measured. The uptake amount of intracellular trehalose is measured before freezing. We find an upper limit for the uptake amount of trehalose when the extracellular trehalose concentration is about 400 mM, at which the freeze-thaw viability is the highest. When the extracellular trehalose concentration exceeds 400 mM, shorter immersion times are needed to obtain the maximum freeze-thaw viability. Also, longer immersion weakens the cells. Our analyses indicate that when the extracellular trehalose-concentration is less than 400 mM, the trehalose uptake occurs more slowly with less dehydration, resulting in less stress on the cell. When the extracellular trehalose concentration exceeds the saturation level, the cell is stressed by the excess dehydration due to the remaining osmotic pressure, with apoptosis occurring before freezing.

show abstract

Section: Freeze-thaw Viabilities Of Cho-vector Cells and Their Apoptosupporting

confidence: 82%

Trehalose uptake and dehydration effects on the cryoprotection of CHO–K1 cells expressing TRET1

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, glycerol showed a good efficacy for phiIPLA-RODI preservation. One possible explanation is that the cryopreservation effect of disaccharides is lost during the slow freezing [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Comparative analysis of different preservation techniques for the storage of Staphylococcus phages aimed for the industrial development of phage-based antimicrobial products

et al. 2018

View full text Add to dashboard Cite

Bacteriophages have been proven as effective antimicrobial agents in the treatment of infectious diseases and in other biocontrol applications including food preservation and disinfection. The extensive use of bacteriophages requires improved methodologies for medium- and long-term storage as well as for easy shipping. To this aim, we have determined the stability of four Staphylococcus phages (phiIPLA88, phiIPLA35, phiIPLA-RODI and phiIPLA-C1C) with antimicrobial potential at different temperatures (20°C/25°C, 4°C, -20°C, -80°C, -196°C) and during lyophilization (freeze drying) using several stabilizing additives (disaccharides, glycerol, sorbitol and skim milk). Differences between phages were observed at different temperatures (20°C/25°C, 4°C and -20°C), where phages were less stable. At lower temperatures (-80°C and -196°C), all phages showed good viability after 24 months regardless of the stabilizer. Differences between phages were also observed after lyophilization although the addition of skim milk yielded a dry powder with a stable titer after 24 months. As an alternative to facilitate storage and transportation, phage encapsulation has been also explored. Phage phiIPLA-RODI encapsulated in alginate capsules retained high viability when stored at 4°C for 6 months and at 20°C for 1 month. Moreover, the spray-dryer technique allowed obtaining dry powders containing viable encapsulated phages (phiIPLA-RODI and phiIPLA88) in both skim milk and trehalose for 12 months at 4°C. Storage of phages at 20°C was less effective; in fact, phiIPLA88 was stable for at least 12 months in trehalose but not in skim milk, while phiIPLA-RODI was stable only for 6 months in either stabilizer. These results suggest that encapsulated phages might be a suitable way for shipping phages.

show abstract

“…This can result in serious cellular damage via increased production of free radical oxygen species [ 18 ]. In addition, recent studies have suggested that trehalose may have an oxidative effect on RBCs via osmotic shock despite its ice crystal inhibiting property [ 19 ]. Thus, optimizing RBCs cold storage and cryopreservation processes to minimize structural and functional damage is an important area of research.…”

Section: Introductionmentioning

confidence: 99%

Salidroside as a Novel Protective Agent to Improve Red Blood Cell Cryopreservation

2016

View full text Add to dashboard Cite

Glycerol and trehalose have been widely examined as protective agents in the cryopreservation of red blood cells (RBCs). However, the effectiveness of these reagents alone on cell viability is moderate. Here, the addition of salidroside attenuated oxidative damage of sheep RBCs prior to and post cryostorage. The supplementation of salidroside to the cryopreservation media containing 10% glycerol improved RBC survival by approximately 61.1±4.8% vs 37.9±4.6%. A smaller effect was seen in RBCs cryopreserved in 300 mM trehalose where the addition of salidroside improved survival by 7.6±0.3%. Furthermore, the addition of salidroside to cold storage solution demonstrated a significant reduction of haemolysis after 4 days for RBCs loaded with either glycerol or trehalose, compared to cells incubated without salidroside. RBCs survival was 2-fold greater following freezing in trehalose, compared with glycerol. After 10 days, salidroside enabled a lower haemolysis of 16.7±1.3% compared to 29.0±8.4% for cells incubated without salidroside. However, salidroside had no effect on RBCs which had been frozen in glycerol as the resulting haemolysis rate by day 10 was approximately 60%. Salidroside increased glutathione reductase activity and decreased lactate dehydrogenase activity. Furthermore, it led to reduced carbonylation of proteins in both glycerol and trehalose loaded cells. Finally, no effect on lipid peroxidation was found in the glycerol loaded RBCs although this was reduced in RBCs loaded with trehalose and salidroside. The present findings confirm the potential use of salidroside as a novel protective agent in cryopreservation and refrigerated storage of sheep RBCs.

show abstract

Freezing Properties of Disaccharide Solutions: Inhibition of Hexagonal Ice Crystal Growth and Formation of Cubic Ice

Cited by 8 publications

References 48 publications

Trehalose uptake and dehydration effects on the cryoprotection of CHO–K1 cells expressing TRET1

Trehalose uptake and dehydration effects on the cryoprotection of CHO–K1 cells expressing TRET1

Comparative analysis of different preservation techniques for the storage of Staphylococcus phages aimed for the industrial development of phage-based antimicrobial products

Salidroside as a Novel Protective Agent to Improve Red Blood Cell Cryopreservation

Contact Info

Product

Resources

About