• Restorative Dentistry and Endodontics
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• 제34권6호
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• pp.491-499
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• 2009

본 연구의 목적은 구강상피세포를 배양한후 각기 다른 조건의 냉동 보존법으로 6일간 보존시 각각의 세포의 활성도를 Cell counting, WST-1, Clonogenic capacity의 방법을 이용하여 비교 평가하기 위함이다. 각 실험군당 $1\times10^6$개의 세포를 다음의 방법으로 6일간 냉동 보존한다. Freezing container에 담아 $1^{\circ}C$/min의 냉동속도로 $-70^{\circ}C$까지 냉동 후 $-196^{\circ}C$에 냉동하여 보관한 일반 냉동 보존군, 세포를 바로 $-196^{\circ}C$의 액화질소에 넣어 냉동한 급속 냉동 보존군, $4^{\circ}C$에서 $-35^{\circ}C$까지 $-0.5^{\circ}C$/min속도로 서서히 냉동시킨 뒤 $-196^{\circ}C$에 냉동한 저속 냉동 보존군, 2 Mpa, 3 Mpa의 압력을 가하고 $-0.5^{\circ}C$/min속도로 $4^{\circ}C$에서 $-35^{\circ}C$까지 서서히 냉동시킨 뒤 $-196^{\circ}C$에 냉동한 2 Mpa, 3 Mpa압력 저속 냉동 보존군으로 나누었다. 6일 후 냉동되었던 세포를 급속 해빙하여 각각의 Cell counting, WST-1, Clonogenic capacity 값을 측정하여 비교하였다. 실험 결과 2 Mpa혹은 3 Mpa의 압력을 이용한 저속 냉동법이 저속 냉동법 및 급속 냉동법 보다 세포 활성도에 있어 우수한 경향을 나타내었다.

• Journal of Microbiology
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• 제39권3호
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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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• 제50권4호
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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.

• 한국식품저장유통학회지
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• 제3권2호
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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.

• 한국식품저장유통학회지
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• 제7권1호
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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.

• 한국수산과학회지
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• 제44권6호
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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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• 제24권6호
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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.

• 한국식품저장유통학회지
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• 제6권1호
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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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• 제35권4호
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• pp.285-294
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• 2010

본 연구의 목적은 흰 쥐의 상악 대구치를 발거한 후 치주인대세포를 $0^{\circ}C$/2 MPa고압-저온하에 1주간 보관시켜 MTT, WST-1 검색법을 이용하여 측정한 치주인대세포의 활성도를 저속 냉동법(No Additional Pressure, 2, 3 MPa), 급속 냉동법(No Additional Pressure, 2 MPa), $-5^{\circ}C$/90 MPa초고압 저온보존법과 비교하여 평가하는 것이다. 생후 4주된 암컷 Sprague-Dawley계 흰쥐의 상악 좌우 제1, 2대구치를 발거하여 각 군 당 12개의 쥐 치아를 MTT, WST-1 검색에 이용하였다. 실험군은 9개군으로 대조군은 즉시 발치군이며, 각각 3 MPa, 2 MPa, No Additional Pressure (NAP)의 압력을 가한 후 $4^{\circ}C$에서 $-35^{\circ}C$까지 $-0.5^{\circ}C$/min 속도로 서서히 냉동시킨 뒤 $-196^{\circ}C$에 냉동한 저속 냉동군, 발치 후 동해방지제 처리과정을 거쳐 각각 2 MPa, NP의 압력을 가한 후, $-196^{\circ}C$의 액화질소에 넣어 냉동한 급속 냉동군, 발치 후 각각 2MPa,NP의 압력을 가한 후, $0^{\circ}C$에 보관한 저온 보존군, $-5^{\circ}C$/90 MPa의 초고압 저온 보존군으로 나누었다. 보존액은 F medium을 사용했으며 동해방지제로 10% dimethylsulfoxide (DMSO)를 사용하였다. 치근면을 단위면적으로 표준화하기 위해 MTT, WST-1 측정값을 Eosin 염색 후 530 nm에서 측정한 흡광도 값으로 나누었다. 통계 분석을 위해 one way ANOVA를 시행하였으며 사후 검정으로는 Tukey HSD 방법을 사용하였고 결과는 다음과 같다. 1. MTT 검색법 및 WST-1 검색법 결과 $0^{\circ}C$/2 MPa 고압 저온 보존군이 즉시 발치군보다 세포 활성도가 낮았으나 통계적 유의차는 없었으며, 저속 압력 냉동군(NP, 2 MPa, 3 MPa)과, 급속 압력 냉동군(NP, 2 MPa), 저온보존군($0^{\circ}C$/NP), 초고압 저온 보존군($-5^{\circ}C$/90 MPa)보다 통계적으로 유의차있게 높은 세포 활성도를 나타내었다(p < 0.05). 2. MTT검색법 및 WST-1 검색법 결과 $-5^{\circ}C$/90 MPa 초고압 저온 보존군이 가장 낮은 세포 활성도를 나타내었으며, MTT 검사 결과에서는 모든 군에 대해 통계적으로 유의성 있는 결과를 보였다(p < 0.05). 위의 결과를 통해, $0^{\circ}C$/2 MPa (20기압)의 고압-저온 보존법이 다른 급속 냉동 보관법(2 MPa, NAP)이나 저속냉동보관법(3, 2 MPa, NAP), $-5^{\circ}C$/90 MPa 초고압 저온 보존법에 비해 우수한 쥐 치아의 치주인대세포의 활성도를 보여 차후 치아의 재이식시 치아보관을 위한 방법으로의 가능성을 제시하였다.

• 한국식품저장유통학회지
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• 제10권4호
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• pp.459-465
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• 2003

본 연구에서는 과채류의 냉동냉장시 내부압력에 따른 조직파괴를 방지하기 위한 기초연구로 동결 방법에 따른 내부압력 변화에 대하여 조사하였다. 과채류의 동결에 의한 중량감소율은 침지식 동결방식에서 0.44∼l.38% 수준으로 가장 적게 나타났으며, 동결방식에 따른 중량감소율 차이는 수박에서 가장 심하여 수분함량에 클수록 동결속도에 따른 효과가 비교적 크다는 것을 알 수 있었다. 동결속도에 따른 내부압력은 배, 사과 및 메론에 있어 침지식 방법에 의한 동결시 체적팽창 및 수축에 따른 내부압력 차가 가장 적게 나타났으며, 송풍식 동결에서 가장 크게 나타나 동결속도가 빠를수록 내압의 크기는 적다는 것을 알 수 있었으나 딸기 및 수박의 경우는 오히려 침지식에서 내압이 가장 크게 나타났다. 균온 처리하지 않은 동결에 있어 과일의 내부압력 변화는 약 2 psig 수준인 반면에 균온 처리한 수박의 내부압력 변화는 균온처리하지 않은 수박의 내부압력 변화 경향과 유사하지만 일정한 수준의 내압을 발생시킴으로써 내부압력의 크기도 약 1.3 psig 수준으로 상당히 적게 나타났다. 또한, 해동시에 있어서도 균온처리한 시료의 내부압력 크기가 균온처리하지 않은 내부압력 크기보다 상대적으로 적게 나타났다. 균온처리 동결시, 다단계 처리보다는 3∼4회 수준의 일정한 균온 처리가 내부압력의 증감 폭을 줄일 수 있었다.