• Journal of Animal Reproduction and Biotechnology
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• v.38 no.1
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• pp.26-31
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• 2023

Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

• Journal of Embryo Transfer
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• v.14 no.3
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• pp.163-169
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• 1999

This study was carried out to investigate the effect of co-culture system(bovine oviduct epithelial cells; BOEC) and defined culture system(modified TALP ; mTALP) on the development of IVM-IVF embryos, and survival of in vitro produced blastocysts after freezing and thawing. Occytes from the slaugheterhous ovaries were matured and fertilized using general protocol. The results obtained were as the following: 1. Survival rates of frozen-thawed blastocysts using 10% glycerol as cryoprotectant was higher in day 7 blastocysts than in Day 8 and 9 blastocysts from co-cultrue system, but survival rate of frozen-thawed blastocysts was higher in Day 10 blastocysts than in day 8 and 9 blastocysts from defined culture system. Regardless of their age, survival rate of frozen-thawed blastocysts was significantly higher (p<0.05) in co-culture system than in defined culture system. 2. The cell number of blastocysts was significanlty higher (p<0.05) in Day 7 blasotcysts than in Day 8 and 9 blastocysts from co-cultures, but the cell number of blsstocysts was significantly higher (p<0.05) in Day 10 blastocysts than in Day 8 and 9 blastocysts from defined culture system. Regardless of the culture system, blastocysts with higher cell number showed higher survival rates after freezing and thawing.

• Journal of Microbiology and Biotechnology
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• v.8 no.5
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• pp.461-465
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• 1998

The suspension culture of an anchorage-dependent cell line (Bok-l) from Bombina orientalis was successful in respects of cost and efficiency. The amount of cells obtained from the suspension culture was almost equivalent to that from the anchorage-dependent culture. This result shows the possibility of suspension culture for scale-up. The cells in suspension produced an antibacterial peptide as much as anchorage-dependent cells did. The cell growth ($6.0\times10^6cells/m\ell$) and viability (>80%) at 10 rpm were higher than that at 0 rpm ($1.9\times10^6cells/m\ell$, 65~80%) and 30 rpm ($1.8\times10^6cells/m\ell$ 40~76%). The size of cells became smaller at the agitation rate of 30 rpm. The antibacterial activities of cell extracts from suspension cultured cells were confirmed against gram-negative and gram-positive bacteria by the inhibition zone assay and the liquid growth inhibition assay.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.894-896
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• 2000

In vitro fertilization can be used for salvaging superior buffalo germplasm which otherwise goes waste after the slaughter of animals. This technology has also increased our basic understanding of growth of germ cells and embryos. The requirement of growing embryos is peculiar and stage specific. In the present study the cleavage and development of buffalo embryos were studied with homologous (buffalo) and heterologous (goat) oviductal cell co-culture systems. The cleavage rate improved significantly (p<0.01) in both homologous and heterologous co-culture as compared to control (55.3, 46.8 and 11.4%). The morula formation using homologous and heterologous oviductal cells also increased significantly as compared to control group (43.6, 21.9 & 1.9%). There was no blastula formation in control group, but addition of oviductal cells either from homologous or heterologous species significantly increased the blastula formation (9.5, 12.5%). The cleavage rate and embryo development was slightly better (non significant) in homologous as compared to heterologous oviductal cell culture. It was concluded that the use of oviductal cell co-culture (homologous and heterologous species) have significantly improved cleavage and development of buffalo embryos in vitro.

• Korean Journal of Environmental Biology
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• v.34 no.2
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• pp.107-115
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• 2016

Currently, there are 442 operating nuclear power plants in the world, and 62 more are under construction. According to this reasoning, the treatment of radioactive waste is important to prevent the environmental ecosystem including humans, animals, and plants. Especially, a leakage of radioactive waste causes not only regional problem but also serious global one. In this study, we demonstrate the effect of radioisotopes (e.g., cesium, strontium, and cobalt) on a 3D culture cell. To develop the 3D cell culture system, we used a 96-well-culture plate with biocompatible agarose hydrogel. Using this method, we can perform the 3D cell culture system with three different cell lines such as HeLa, HepG2, and COS-7. In addition, we conducted a cell viability test in the presence of radioisotopes. Interestingly, the 3D morphological cells showed 42% higher cell viability than those on the 2D against cesium. This result indicates that the 3D platform provides cells morphological and physiological characteristic similar to in vivo grown tissues. Moreover, it overcomes the limitation of conventional cell culture system that can't reflect in vivo systems. Finally, we believe that the proposed approach can be applied a new strategy for simple high-throughput screening and accurate evaluation of metal toxicity assay.

• Korean Journal of Agricultural Science
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• v.20 no.1
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• pp.34-42
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• 1993

Effects of salt concentrations on the cell growth and the pigment accumulation were investigated in cell suspension culture of Vitis vinifera and Phytolacca americana L.. The growth pattern of vine cell in control was showed the normal exponential growth pattern, but in the dilution media delay the exponential growth pattern from 4 to 8 days after culture. Maximal accumulation of anthocyanin was observed at 12 days after culture in all treatments. In cell suspension culture of Phytolacca, accumulation of betacyanin occurred in parallel with the cell growth pattern and maximal accumulation of betacyanin was observed after 8 days of culture. In the vine cell culture, the cell growth was showed the peak at 87.6mM of sucrose in the medium and reduced at over this concentration. Maximal anthocyanin accumulation was showed at 146mM of sucrose. In the higher concentrations of sucrose, the cell growth was rapidly decreased, but the accumulation of anthocyanin was not. Otherwise, in case of Phytolacca cell culture, betacyanin accumulation was showed in parallel with the cell growth increased with sucrose concentration. It was suggested that the anthocyanin of vine and the betacyanin of Phytolacca were controlled by different mechanisms.

• Journal of Microbiology
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• v.42 no.3
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• pp.248-252
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• 2004

In order to identify the protein(s) secreted into culture medium by the sool-l/retl-l mutation of Saccharomyces cerevisiae, proteins from the culture medium of cells grown at permissive (28$^{\circ}C$) and non-permissive temperatures (37$^{\circ}C$), were analyzed. Comparison of protein bands separated by SDS-PAGE identified a prominent band of 47-kDa band from a mutant grown at 37$^{\circ}C$. N-terminal amino acid sequencing of this 47-kDa protein showed high identity with enolases 1 and 2. Western blot analysis revealed that most of the cell wall-bound enolase was released into the culture medium of the mutant grown at 37$^{\circ}C$, some of which were separated as those with lower molecular weights. Our results, presented here, indicate the impairment of cell wall enolase biogenesis and assembly by the sool-l/retl-l mutation of S. cerevisiae.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.512-519
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• 1997

RNA accumulating strain of Torulopsis versatilis MT-1 was cultured in molasses medium for higher contents of RNA in cell. Yeast cells were harvested at logarithmic phase on synchronous culture. Yield of cells on dry base to input sugar was 59.5%. Crude protein content was 55.1% in cell. RNA content was 13.9%. Some problems found in the process for the preparation of yeast extracts were improved by the addition of culture broth of Streptomyces faecalis MSF which secrete RNase (5' nuclease and 5' adenylic acid deaminase). When the culture broth of S. faecalis MSF was added in autolysis process 46% of RNA in cell was converted to I and G(5' inosinic acid and 5' guanylic acid) in extract. By addition of 3-7% culture broth of S.faecalis MSF in autolysis or enzymolysis process at the start or early stage, RNA in extract was converted easily to I and G and protein in cells was easily extracted and hydrolyzed to amino acid. Taste of those yeast extracts was delicious. The yeasty smell in yeast extracts was removed. And cell debris was easily removed from extract.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.312-315
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• 2005

A process for efficient recycle fed-batch culture was carried out to increase cell mass and spore production by Lactic acid bacteria isolated from Kimchi. A large quantity of cell mass obtained by feeding concentration of sugar in recycle fed-batch culture. When the high density of salt was created that the cell mass was come-down. In this study, cultured in different feeding concentration of sugar conditions. Lactic acid bacteria by recycle fed-batch culture was investigated in 2L working volume of fermenter, obtained the maximum cell mass was 15.17g/L.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.124-128
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• 2010

Vaccines, especially for viruses, have been produced from egg-based manufacturing process. The method is simple and easy to set up the manufacturing process. However, the method has many problems in quality control, limit of manufacturing capacity, and ethical issues. Over the last decade, an alternative method, which manufactures vaccines using cell culture-based system, has received great attention to overcome the problems in egg-based vaccine production. This article examines current status and perspectives of cell culture-based vaccine production.