• Restorative Dentistry and Endodontics
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• v.34 no.6
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• pp.491-499
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• 2009

This study examined the influence of the storage methods on the viability of oral epithelial cells using conventional cell freezing storage, slow freezing preservation, rapid freezing preservation, and slow freezing preservation with a pressure of 2 Mpa or 3 Mpa. The cell viability was evaluated by cell counting, WST-1 and the clonogenic capacity after 6 days of freezing storage. After 6 days, the frozen cells were thawed rapidly, and the cell counting. WST-1, and clonogenic capacity values were measured and compared. 1. The results from cell counting demonstrated that conventional cryopreservation, slow freezing under a 2 Mpa pressure and slow freezing under a 3 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p < 0.05). 2. The results from the optical density by WST-1 demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 3. The clonogenic capacity demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p < 0.05).

• Journal of Microbiology
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• v.39 no.3
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• pp.240-243
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• 2001

The effect of cryoprotectants and suspending solutions on the preservation of marine heterotophic bacteria was investigated. Six halotolerant and four halophilic bacterial isolates suspended in either distilled water or artificial seawater were preserved in glycerol and dimethylsulfoxide at -70$\^{C}$, respectively. After one year of preservation, the recovery rates on the appropriate agar plates were estimated. The survival rate was found to be dependent on the strain tested, regardless of the preservation conditions tested.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.213-222
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• 2023

Cryopreservation is an option for the preservation of pre- or post-pubertal female or male fertility. This technique not only is beneficial for human clinical applications, but also plays a crucial role in the breeding of livestock and endangered species. Unfortunately, frozen germ cells, including oocytes, sperm, embryos, and spermatogonial stem cells, are subject to cryoinjury. As a result, various cryoprotective agents and freezing techniques have been developed to mitigate this damage. Despite extensive research aimed at reducing apoptotic cell death during freezing, a low survival rate and impaired cell function are still observed after freeze-thawing. In recent decades, several cell death pathways other than apoptosis have been identified. However, the relationship between these pathways and cryoinjury is not yet fully understood, although necroptosis and autophagy appear to be linked to cryoinjury. Therefore, gaining a deeper understanding of the molecular mechanisms of cryoinjury could aid in the development of new strategies to enhance the effectiveness of the freezing of reproductive tissues. In this review, we focus on the pathways through which cryoinjury leads to cell death and propose novel approaches to enhance freezing efficacy based on signaling molecules.

• Food Science and Preservation
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• v.3 no.2
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• pp.121-129
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• 1996

Plum highlighted as a health food is needed to diversify the processed products because labor storage is big problem since the fruit was producted massively in June. The Plum was extracted by the pressing type extractor after washing, drying and removing the seed by seed separator. The crude extract was concentrated with stainless steel vessel at different temperature and stirring speed. This study was obtained as follows. The sugar content of fresh plum concentrated extract was 55.3~58.3$^{\circ}$Brix, and of the freezing plum concentrated extract was 75.5~70.3$^{\circ}$Brix. In color difference, the freezing plum concentrated extract was more deep black than fresh plum. In change patten of pH, it was decreased as concentration was proceed. The final pH was 2.3~2.2 in fresh plum, and 1.8~2.2 in freezing plum. The total acid content of fresh plum concentrated extract and the freezing plum was 45.4~47.8, 60.3~60.9%, respectively. The content of evaporation at 85$\pm$5$^{\circ}C$ was constant irrespective of stirring speed. The yield of extraction of fresh plum was higher than freezing plum. According to this results, the use of stainless vessel, 50rpm, which gave a highly qualified plum concentrated extract.

• Food Science and Preservation
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• v.7 no.1
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• pp.57-62
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• 2000

Ginseng was stored at a temperature lower than the freezing point after a treatment of freezing point depressing agents to extend its freshness. Respiration rate at freezing point of ginseng, -1.7${\pm}$0.1$^{\circ}C$, was inhibited 92% and 97% compared with those stored at 5$^{\circ}C$or 20$^{\circ}C$ , respectively. Sorbitol solution chosen as a freezing point depressing agent lowered the freezing point of ginseng to about -3.0$^{\circ}C$. Ginsengs treated with the sorbitol solution and packaged with 0.06mm LDPE was stored at -2$^{\circ}C$ , and the quality change was then compared with ginsengs stored at 0$^{\circ}C$ and 5$^{\circ}C$. Weight loss of ginsengs stored at -2$^{\circ}C$ for 100days was 1.5%, which is about 2.6times less than those stored at 5$^{\circ}C$. However, there were no significant difference between the ginsengs stored at -2$^{\circ}C$ and at 0$^{\circ}C$(1.9%). Spoilage rate of the ginsengs was 100% after 50 days of storage at 5$^{\circ}C$ and 25% after 100days at 0$^{\circ}C$respectively. but that of ginsengs stored at -2$^{\circ}C$ was 13%, which was half than that of ginsengs stored at 0$^{\circ}C$. Firmness and amount of monoscaccharides in ginsengs were decreased during storage at 5 or 0$^{\circ}C$ but ginsengs stored at -2$^{\circ}C$ showed better firmness and an increase in monosaccharides such as fructose and glucose. From above, when ginseng treated with freezing pont depressing agents were stored at -2$^{\circ}C$, the shelf life was extended to 2 or 3 times longer than those that were stored at 5 or 0$^{\circ}C$, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.6
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• pp.665-670
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• 2011

The rotifer Brachionus plicatilis is one of the most important food organisms in aquaculture. The resting eggs produced by mictic female rotifers are easily stored and hatched, making them useful as the starter for the mass culture of rotifers in marine larval culture. This study examined the optimum preservation method for resting eggs to ensure a high hatching rate. To produce resting eggs, the marine rotifer B. plicatilis was cultured with Nannochloris oculata (KMMCC 16). The resting eggs were harvested and cryopreserved using 5% and 10% methanol (MeOH), dimethylsulfoxide (DMSO), and glycerol as cryoprotectant agents (CPAs). The cryopreservation comprised slow or rapid freezing and the resting eggs were stored for one month in liquid nitrogen ($-196^{\circ}C$). The resting eggs were also dried at different temperatures (30, 40, and $50^{\circ}C$) and for different times (1, 2, and 3 h). In general, the hatching rates of the resting eggs preserved with CPA were higher than those without CPA and the slow freezing method was better than the rapid freezing method. However, the optimum CPA concentration for the hatching rate of the resting eggs varied with the freezing method and kind of CPA, and the CPA also affected the viability of the resting eggs. Dried resting eggs had a high, rapid hatching rate over 80%. The moisture content of the resting eggs cryopreserved in liquid nitrogen affected the hatching rate. Drying at $30^{\circ}C$ for 1 hour resulted in a high hatching rate of the resting eggs. In conclusion, drying at $30^{\circ}C$ for 1 hour and preservation in liquid nitrogen with the slow freezing method, without CPA, is recommended for a high hatching rate (ca. 95%) of rotifer resting eggs.

• Journal of Veterinary Science
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• v.24 no.6
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• pp.79.1-79.16
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• 2023

Background: The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively recently introduced preservation method that meets this need. Supercooling refers to the phenomenon of liquid in which the temperature drops below its freezing point without solidifying or crystallizing. Objectives: The purpose of this study was to identify the preservation efficiency and applicability of the supercooling technique as a cortical bone allograft storage modality. Methods: The biomechanical effects of various storage methods, including deep freezing, cryopreservation, lyophilization, glycerol preservation, and supercooling, were evaluated with the three-point banding test, axial compression test, and electron microscopy. Additionally, cortical bone allografts were applied to the radial bone defect in New Zealand White rabbits to determine the biological effects. The degree of bone union was assessed with postoperative clinical signs, radiography, micro-computed tomography, and biomechanical analysis. Results: The biomechanical properties of cortical bone grafts preserved using glycerol and supercooling method were found to be comparable to those of normal bone while also significantly stronger than deep-frozen, cryopreserved, and lyophilized bone grafts. Preclinical research performed in rabbit radial defect models revealed that supercooled and glycerol-preserved bone allografts exhibited significantly better bone union than other groups. Conclusions: Considering the biomechanical and biological superiority, the supercooling technique could be one of the optimal preservation methods for cortical bone allografts. This study will form the basis for a novel application of supercooling as a bone material preservation technique.

• Food Science and Preservation
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• v.6 no.1
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• pp.81-86
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• 1999

The textural characteristics of nonxaxy rice flour gels and rice cake(Injolmi) with different water contents and additives were evaluated after freezing and microwave heating. As moisture content of rice flour gels increased from 45% to 55%, its hardness and gumminess decreased, but adhesive and cohesiveness had no significant difference. Microwave heating did not markedly affect the texture but frozen storage was very effective to prevent the hardening of products. Hardness of reheated rice gels increased more rapidly in non-packaged sample than in PE wrap film and affected by storage time of 24hrs at 20$^{\circ}C$. As sugar content of rice flour gels increased from 0% to 10%, its hardness, adhesiveness, and gumminess decreased, while cohesiveness did not change.

• Restorative Dentistry and Endodontics
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• v.35 no.4
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• pp.285-294
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• 2010

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa), group 8 (low-temperature preservation at $0^{\circ}C$ under no additional pressure), group 9 (low-temperature preservation at $-5^{\circ}C$ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 ($0^{\circ}C$/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.

• Food Science and Preservation
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• v.10 no.4
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• pp.459-465
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• 2003

This study was carried out to investigate the changes in internal pressure according to various freezing methods, as basic research to protect the destruction of tissues when fruits and vegetables are frozen. The rate of weight loss, caused by the freezing of fruits and vegetables, was found to be the least (0.44∼1.38％) when the immersion freezing method was applied. The difference in the rate of weight loss was the highest when freezing methods were applied to watermelon, and the freezing rate of watermelon whose moisture contents were greater have relatively greater influence on the weight loss. The difference in internal pressures was the least and caused by the volume increase and decrease, when pear, apple, and melon were frozen using the immersion freezing method, while the diffeyence the greatest when the air-blast freezing method was used. As the freezing rate was greater, the internal pressure was less. However, the internal pressure of strawberry and watermelon was the greatest when the immersion freezing method was applied. Frozen without using the thermal equalizing method, the change in internal pressure of fruits was about 2 psig. In contrast, the internal pressure of watermelon applied with the thermal equalizing method was changed in a way similar to that of watermelon not applied with the method, but the former generated a certain level of internal pressure and maintained a significantly low level of internal pressure (about 1.3 psig). When thawed, the internal pressure of samples to which the thermal equalizing method was applied was less than that of what the thermal equalizing method was not applied to. In comparison with the application of multi-step thermal equalizing method, 3∼4 times of application of the thermal equalizing method to the freezing resulted in the decrease of fluctuation range of internal pressure.