• KSBB Journal
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• 제21권4호
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• pp.306-311
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• 2006

세포의 보존은 세포의 배양에 있어서 필수 불가결한 요건으로서 세포주의 다양한 특성에 따라 적합한 방법이 확립되어야 한다. 본 연구에서는 산업용 세포주인 CHO 세포의 단기간 저온보존 기술의 확립을 목표로 진행되었으며 다양한 조건을 통해 가장 안정적인 저온보존 방법을 수립하였다. 저온보존 방법에 있어서 가장 중요한 요인은 온도로서 $4^{\circ}C$ 저온보존이 세포 보존에 필수적인 조건으로 나타났으며 $20^{\circ}C$ 실온보존에서는 세포의 급격한 사멸이 관찰되었다. 보존형태는 용기를 눕힌 상태로 서서히 회전시켜 현탁 보존하는 방법이 용기를 세우거나 눕혀 보관하는 방법에 비해 높은 생존율을 나타내었다. 또한 저온보존 시 새로운 배지로 교환한 후 보존하는 방법이 배양에 사용된 배지를 그대로 사용한 보존 방법보다 세포의 성장 회복율에서 우수한 것으로 나타났다. 하지만 $4^{\circ}C$에서 rolling을 통한 현탁 보존을 할 경우에는 배지의 교환 없이도 안정적으로 세포보존이 가능한 것으로 나타났다. 저온보존에 가장 적합한 세포의 농도는 실험결과 $1.0{\times}10^6{\sim}5.0{\times}10^6cells/m{\ell}$ 범위로 나타났으며 혐기적인 상태로 보존하는 것이 공기가 존재하는 보존방법 보다 비교적 우수한 보존 결과를 나타내었다. 이상의 결과를 바탕으로 무혈청 배지의 저온보존액으로서의 안정성과 첨가물에 의한 보존효율의 향상을 평가하였다. 실험결과 저온보존 후 10일간은 높은 세포 생존율과 함께 정상적인 세포 성장 회복을 보이는 것으로 나타났으며 ${\alpha}$-tocopherol과 retinoic acid를 첨가한 저온보존액의 경우에는 더욱 우수한 세포 생존율을 보임을 확인하였다. 마지막으로 이렇게 확립된 방법을 이용하여 1 L 용량의 저온보존 실험을 수행한 결과, 앞선 실험에서와 유사한 경향의 세포 보존 능력을 확인할 수 있었다. 이러한 결과를 종합해 볼 때 산업용 세포주로 널리 사용되는 CHO 세포의 저온보존은 본 연구에서 확립된 방법을 통해 단기간 동안 안정적으로 수행될 수 있을 것으로 사료되며 대용량 저온보존의 적용 가능성도 확인하였다. 대용량 배양에서의 단기간 보존기술에 대한 연구가 앞으로 더 많이 수행된다면 실제 배양 공정에서도 저온보존 기술의 적용이 가능할 것으로 판단된다.

• 한국물환경학회지
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• 제34권4호
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• pp.375-381
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• 2018

The monitoring of phytoplankton biomass and community structure is essential as a first step to control the harmful cyanobacterial blooms in freshwater systems, such as seen in rivers and lakes, due to the process of eutrophication and climate change. In order to quantify the biomass of phytoplankton with a wide range in size and shape, the measurement of cell biovolume along with cell density is required for a comprehensive review on this issue. However, most routine monitoring programs preserve the gathered phytoplankton samples before analysis using chemical additives, because of the constraint of time and the number of samples. The purpose of this study was to investigate the cell biovolume change characteristics of six cyanobacterial species, which are common bloom-causing cyanobacteria in the Nakdong River, after the preservation with Lugol's iodine solution. All species showed a statistically significant difference after the addition of Lugol's iodine solution compared to the live cell biovolume, and the cell biovolume decreased to the level of 34.0 ~ 56.3 % at maximum in each species after the preservation. The nonlinear regression models for determining the shrinkage ratio by a preservation period were derived by using the cell biovolume measured until 180 days preservation of each target species, and the equation to convert the cell biovolume measured after preservation for a certain period to the cell biovolume of viable cell was derived using that formula. The conversion equation derived from this study can be used to estimate the actual cell biovolume in the natural environment at the time of sampling, by using the measured biovolume after the preservation in the phytoplankton monitoring. Moreover this is expected to contribute to the final interpretation of the water quality and aquatic ecosystem impacts due to the cyanobacterial blooms.

• KSBB Journal
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• 제24권3호
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• pp.312-320
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• 2009

치료용 단백질을 생산하는 생물의약품 산업이나 세포치료제 및 이식용 세포를 다루는 재생의학 분야 등의 세포기반 산업에서 안정적인 세포의 보존은 필수적인 요소이다. 본 연구에서는 인간 피부 섬 유아세포의 $4^{\circ}C$ 저온보존에서 우수한 성능을 나타내는 개선된 저온보존액을 개발하고, 저온에 의한 세포 손상을 보호함으로써 보다 안정적인 세포 저온보존 기술을 제공하고자 하였다. 세포의 저온보존에서 우수한 효능을 나타내는 핵심 성분을 탐색한 결과, yeast hydrolysate 등의 단백질 가수분해물을 첨가한 보존액에서 월등히 뛰어난 보존효과가 나타남을 확인하였다. 단백질 가수분해 물은 미생물, 식물, 동물유래 단백질 가수분해 물에서 모두 우수한 효과를 나타냈으며, 특히 단백질 가수분해물 성분 중 분자량 10kDa 이하의 펩타이드를 첨가한 저온보존에서 우수한 보존효과가 나타났다. 저온에 의한 세포손상에 대해 단백질 가수분해물은 세포내 ATP level의 감소를 막아주고 ROS 생성을 억제하는 것으로 나타났으며, 항산화제 및 삼투압 조절물질을 단백질 가수분해 물과 함께 첨가하였을 때 더욱 우수한 세포 보존효과를 보였다. 최종적으로 본 연구에서 개발한 KUL261 저은보존액 (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%)은 기존의 저온 보존액에 비해 세포 생존을 및 성장률에서 월등히 우수한 성능을 나타내었다. 결론적으로, 핵심 유효성분으로 단백질 가수분해물을 포함하는 개선된 저온보존액은 기존의 보존액보다 월등히 우수한 보존효과를 제공하며, 세포치료제 및 재생의학 분야의 발전과 글로벌 상업화에 기여할 수 있을 것이다.

• 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.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.170.2-170.2
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• 2006

• 한국멀티미디어학회논문지
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• 제24권10호
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• pp.1326-1335
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• 2021

As part of the cell division method, we proposed a method for segmenting images generated by topography microscopes through deep learning-based feature generation and graph segmentation. Hybrid vector shapes preserve the overall shape and boundary information of cells, so most cell shapes can be captured without any post-processing burden. NIH-3T3 and Hela-S3 cells have satisfactory results in cell description preservation. Compared to other deep learning methods, the proposed cell image segmentation method does not require postprocessing. It is also effective in preserving the overall morphology of cells and has shown better results in terms of cell boundary preservation.

• 한국식품저장유통학회지
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• 제1권2호
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• pp.107-116
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• 1994

This paper was carried out to Investigate changes in the activities of cell-degrading enzymes, cell wall components and cell structure of peach during maturation and storage for valuation of quality. The firmness of peach was decreased during maturation and storage, and was remarkably decreased in Daegubo than Yumyung. Polygalacturonase and $\beta$-galactosidase activities of peach were increased during maturation and storage, and were remarably increased in soft peach and in mature and soft peach, respectively. Contents of alcohol-insoluble substance, cell wall, and total and insoluble pectin of peach were decreased during maturation and storage, but cellulose and soluble pectin were increased. Intracellular space was enlarged during maturation and middle lamella was gradually degraded during maturation.

• Reproductive and Developmental Biology
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• 제38권1호
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• pp.1-8
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• 2014

Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxygen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권1호
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• pp.41-46
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• 2003

Difficulties associated with bioartificial liver (BAL) preservation limit not only the commercialization of BAL, but also its clinical trials. In this study, the possibility of cold preservation of BAL cartridges containing porcine hepatocytes was examined at 4$^{\circ}C$. In an in vitro perfusion culture System, BAL cartridges maintained cytochrome P450 metabolic function for at least 50 days. However, all BAL cartridges completely lost their ammonia eliminating ability when stored at 4$^{\circ}C$. We a1so studied the effect of cell density on the maintenance of BAL liver function In a highly differentiated and healthy state. As expected, BALs containing a larger number of hepatocytes demonstrated higher metabolic functions. When metabolic functions were compared per gram of hepatotytes, no large differences were observed between devices containing different densities of hepatocytes. Decreased cell density did not Successfully prolong BAL function. The viability and function of isolated hepatotytes highly depend on the culture conditions, such as cell density, substrata, culture media, and additives to the culture media. Perfusion culture of BAL cartridges at 4$^{\circ}C$ gave a promosing result with respect to the maintenance of P450 activity. However, as indicated by the rapid loss of ammonia metabolic activity, many factors still remain to be optimized for preservation of BAL keeping high metabolic functions for a longer time.

• Nutrition Research and Practice
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• 제1권4호
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• pp.260-265
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• 2007

Preservation methods on the physiological and brewing technical characters in bottom and top brewing yeast strains were investigated. The preserved yeasts were reactivated after 24 months storage and grown up to stationary phase. The samples of filter paper storage indicated a higher cell growth and viability during propagation than those of nitrogen and lyophilization storage independent on propagation temperature. In addition, the filter paper storage demonstrated a faster absorption of free amino nitrogen and a highest level of higher aliphatic alcohols production during propagation than other preservation methods, which can be attributed to intensive cell growth during propagation. Moreover, the filter paper storage showed a faster accumulation for glycogen and trehalose during propagation, whereas, in particular, lyophilization storage noted a longer adaptation time regarding synthesis of glycogen and trehalose with delayed cell growth. In beer analysis, the filter paper storage formed an increased higher aliphatic alcohols than control. In conclusion, the preservation of filter paper affected positively on yeast growth, viability and beer quality independent on propagation temperature. In addition, in this study, it was obtained that the HICF and Helm-test can be involved as rapid methods for determination of flocculation capacity.