• Title/Summary/Keyword: Cell preservation

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Short-term Hypothermic Preservation of CHO Cells Using Serum-Free Media (무혈청 배지를 이용한 CHO 세포의 단기 저온보존)

  • Byoun, Soon-Hwi;Park, Hong-Woo;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.4
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    • pp.306-311
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    • 2006
  • Cell preservation is indispensable in animal cell culture process and should be established according to the cell characteristics. In this study, we experimented hypothermic preservation of CHO cells that is widely used in pharmaceutical industry to produce therapeutic proteins and established a stable method of preservation. The highest viability of CHO cells was obtained when the cells were preserved using rolling tube, which means the cells should be suspended to avoid the cell lumping during the preservation. Also, we obtained superior preservation result under the anaerobic condition. To evaluate the serum-free media as a preservation solution, we investigated cell growth after hypothermic preservation using serum-free media. High cell viability and normal cell growth was observed during 10 days using serum-free media. Moreover, we found that more effective preservation when ${\alpha}$-tocopherol and retinoic acid is added to media as an additive. In the case of 1 liter large scale hypothermic preservation using established protocol, cell viability and growth rate was obtained as good as small scale one. This study is considered to be helpful for hypothermic preservation of CHO cells and large scale hypothermic preservation may be available through the further studies.

Effect of Lugol's Iodine Preservation on Cyanobacterial Biovolume and Estimate of Live Cell Biovolume Using Shrinkage Ratio (Lugol's Iodine Solution 첨가 후 보존 기간별 남조류 세포부피 변화 및 수축비를 이용한 생세포 부피 산정)

  • Park, Hae-Kyung;Lee, Hyeon-Je;Lee, Hae-Jin;Shin, Ra-Young
    • Journal of Korean Society on Water Environment
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    • v.34 no.4
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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.

Development of hypothermic preservation solution for the human dermal fibroblast using protein hydrolysates (단백질 가수분해 물을 이용한 인간 피부 섬유아세포의 저온 보존액 개발)

  • Byoun, Soon-Hwi;Choe, Tae-Boo
    • KSBB Journal
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    • v.24 no.3
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    • pp.312-320
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    • 2009
  • Stable cell preservation is an essential factor in the regenerative medicine for cell therapies and transplantation of biologic materials. In this study, we studied to provide more stable hypothermic preservation by protection of cell damage during the preservation at $4^{\circ}C$. The result of searching for key components that have excellent efficacy in hypothermic preservation of cells, we have identified the fact that the hypothermic preservation adding protein hydrolysates such as yeast hydrolysate is far superior to others. All protein hydrolysates that are derived from animal, plant and microbe sources have superior efficacy, especially the peptides which have molecular weights under 10 kDa have the best efficacy among the components of protein hydrolysate. The protein hydrolysates prevented the decrease of ATP level in the cells caused by hypothermic environment and they inhibited the generation of ROS. Adding antioxidants and control agents of osmotic pressure were showed to have more superior efficacy in hypothermic preservation. Finally, KUL261 solution (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%), the preservation solution developed in this study, showed the best efficacy in both cell viability and cell growth more than other conventional preservation solutions. In conclusion, the improved hypothermic preservation solution that contains the protein hydrolysates as a key component provide the best preservation efficacy. It provides better efficacy than other preservation solutions and will contribute to both the development of regenerative medicine and global commercialization in this therapeutic field.

COMPARISON OF VIABILITY OF ORAL EPITHELIAL CELLS STORED BY DIFFERENT FREEZING METHODS (구강상피세포의 냉동보관 방법에 따른 세포생존률 비교)

  • Baek, Do-Young;Lee, Seung-Jong;Jung, Han-Sung;Kim, Eui-Seong
    • 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).

Bio-Cell Image Segmentation based on Deep Learning using Denoising Autoencoder and Graph Cuts (디노이징 오토인코더와 그래프 컷을 이용한 딥러닝 기반 바이오-셀 영상 분할)

  • Lim, Seon-Ja;Vununu, Caleb;Kwon, Oh-Heum;Lee, Suk-Hwan;Kwon, Ki-Ryoug
    • Journal of Korea Multimedia Society
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    • v.24 no.10
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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.

Changes in the Cell Wall Components, Enzyme Activities and Cell Structure of Peach during Maturation (복숭아의 품종과 숙도에 따른 세포벽 성분, 효소활성 및 조직의 변화)

  • 김미현;신승렬
    • Food Science and Preservation
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    • v.1 no.2
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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.

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Storage of Bull and Boar Semen: Novel Concepts Derived Using Magnetized Water and Antioxidants

  • Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.38 no.1
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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.

Effect of Low Temperature Preservation and Cell Density on Metabolic Function in a Bioartificial Live

  • Park, Yueng-Guen;Takehiko Tosha;Satoshi Fujita;Boru Zhu;Hiroo Iwata;Ryu, Hwa-Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.1
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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.

Influence of preserved brewing yeast strains on fermentation behavior and flocculation capacity

  • Cheong, Chul;Wackerbauer, Karl;Beckmann, Martin;Kang, Soon-Ah
    • Nutrition Research and Practice
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    • v.1 no.4
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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.