• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.349-357
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• 2021

Monoclonal antibodies are widely used as diagnostic reagents and for therapeutic purposes, and their demand is increasing extensively. To produce these proteins in sufficient quantities for commercial use, it is necessary to raise the output by scaling up the production processes. This review describes recent trends in high-density cell culture systems established for monoclonal antibody production that are excellent methods to scale up from the lab-scale cell culture. Among the reactors, hollow fiber bioreactors contribute to a major part of high-density cell culture as they can provide a tremendous amount of surface area in a small volume for cell growth. As an alternative to hollow fiber reactors, a novel disposable bioreactor has been developed, which consists of a polymer-based supermacroporous material, cryogel, as a matrix for cell growth. Packed bed systems and disposable wave bioreactors have also been introduced for high cell density culture. These developments in high-density cell culture systems have led to the monoclonal antibody production in an economically favourable manner and made monoclonal antibodies one of the dominant therapeutic and diagnostic proteins in biopharmaceutical industry.

• Journal of Microbiology and Biotechnology
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• v.1 no.2
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• pp.126-129
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• 1991

In order to produce ${\gamma}-linolenic$ acid by Mucor sp. KCTC 8405P. the fungus was cultivated in fed-batch culture with two phases. i.e., growth in yeast-like form and induction to hyphal growth by pH shift of the culture medium during cultivation. The synchronous growth of the fungus into the appropriate sizes was important for the high density cell culture of this dimorphic fungus. Dissolved oxygen concentration in the medium did not affect degree of unsaturation of fatty acids and ${\gamma}-linolenic$ acid content. Under the culture conditions applied in this experiment. the fungus was found to produce 100 g/l dry mycelia containing 40% of the lipids, where ${\gamma}-linolenic$ acid comprised about 9% of the total extractable fatty acids.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.753-757
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• 1999

The growth of rotifer, Brachionus rotundiformis was evaluated at different culture conditions. Rotifer was fed on condensed freshwater Chlorella. The productivity of rotifer in the high density culture system was compared to that of rotifer in the batch culture system, in which rotifer was fed on baker's yeast. The growth rate of rotifer increased as temperature increased in the culture system supplied with air or oxygen gas. The maximum density of rotifer in the culture systems supplied with air was in range of 16,300$\~$17,000 ind./ml at $24^{\circ}C$. In the culture systems supplied with oxygen gas, it ranged 26,300$\~$30,500 ind/ml at $28^{\circ}C$. When the concentration of dissolved oxygen in the culture system supplied with air reached to below 1 ppm or when the concentration of undissolved ammonia in the culture system supplied with oxygen gas reached 16.6$\~$22.6 ppm, the growth of rotifer decreased. When oxygen gas was supplied and pH was adjusted to 7, the maximum density of rotifer reached to 43,000 ind/ml at $32^{\circ}C$. The production costs for 10 billion rotifer in the high density culture and batch culture were 693,000 and 961,000 won, respectively. Therefore, it was concluded that the productivity of rotifer in the high density culture was higher than that in a batch culture.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.123-129
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• 2000

A high cell density culture system for the anchorage dependent CHO cells was developed based on the combination of in removal of ammonium ion and microcarrier culture system, and semi-fed-batch feeding of glucose and glutamine was employed to the developed culture system. The glass bead was selected as an optimum microcarrier in terms of cell growth. An ammonium ion selective zeolite, Phillipsite-Gismondine, was packed in a dialysis menium ion. The semi-fed-batch operation was employer to the novel culture system for the high density cell culture, and the results showed the cell growth was improved by 32% and tPA productivity by 250%.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.88-88
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• 2017

The study was conducted to analyze the growth characteristics comparison per planting density on the waxy corn early-planting culture for the paddy field in the southern province of south Korea. The cultivation period of early-planting culture for the paddy farming of the waxy corn are sown on February 15, 2016years, transplanting March 15 and harvest June 20. And it grew 126 days. The weather change according to the cultivation period of unheated plastic house early-planting culture, it was average temperature $14.6^{\circ}C$ and humidity 62.5%. And the temperature was 5.6 degrees Celsius warmer compared with the outside temperature and the humidity was 0.7 percent higher tendency. At the growth per planting density of waxy corn, culm length was average 224cm, the more it is high density culture the more was high trend. Stem diameter and ear length the more it is high density culture the more was lowed trend. The node number of $60{\times}20Cm$ was 12 nodes, fruit seting 5.7 nodes, tasseling number 94 days and silking number 96 days. In the ear characteristics per planting density, the size of ear length, seed setting length, ear width and ear weight the more planting density is high the more lowed that trend. The commodity percentage of planting density $60{\times}35Cm$ was the highest among other treatment as 69.1%. But, marketable yield was the highest planting density of $60{\times}20Cm$ as 4,543 ears/10a, and appeared in order $60{\times}25Cm$ 95%> $60{\times}30Cm$ 93%> $60{\times}35Cm$ 92%. The planting density on the waxy corn early-planting culture for the paddy farming in the southern province, the planting density analyzed to be effective planting of over 25% than normal season culture.

• ALGAE
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• v.38 no.4
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• pp.283-294
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• 2023

Previous research indicated that free-living sporangial filament keep hollow morph under high-culture density and form bipartite cells under low-culture density, while the following conchospore release was inhibited by high light. Here, we further explored the molecular bases of these affects caused by light and culture density using a transcriptome analysis. Many differentially expressed genes (DEGs) related to carbon dioxide concentration and fixation, photosynthesis, chlorophyll synthesis and nitrogen absorption were upregulated under high-light conditions compared with low-light conditions, indicating the molecular basis of rapid vegetative growth under the former. The stress response- and ion transport-related DEGs, as well as the gene encoding the vacuole formation-brefeldin A-inhibited guanine nucleotide exchange protein (BIG, py05721), were highly expressed under high-density conditions, indicating the molecular basis of the hollow morph of free-living sporangial filaments under high-culture density conditions. Additionally, the brefeldin A treatment indicated that the hollow morph was directly influenced by vacuole formation-related vesicle traffic. Others DEGs related to cell wall components, zinc-finger proteins, ASPO1527, cell cycle and cytoskeleton were highly expressed in the low density with low-light group, which might be related to the formation and release of conchospores. These results provide a deeper understanding of sporangial filaments in Neopyropia yezoensis and related species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.2
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• pp.93-97
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• 2000

This study was carried out to investigate the effect of rotifer fed the different diets in high density culture on larval flounder Paralichthys olivaceus. Potifer was enriched with enrichment supplements, Marine ${\alpha}\;and\;{\omega}-yeast$ for 6 hours after being cultured with freshwater Chlorella for 18 hours during high density culture before it was fed to larval flounder. And rotifer was culutured with marine Chlorella and freshwater Chlorella for 24 hours during semi-continuous high density culture before it was fed to larval flounder. Culture tanks(21 working volum) set for rotifer culture in a water bath($28{\circ}C$) were continuously supplied with oxygen gas. The content of n-3 HUFA to fatty acids in rotifer(dry weight ${\%}$) enriched with Marine ${\alpha}$ for 6 hours and cultures with marine Chlorella for 24 hours were higher than that in rotifer enriched with ${\omega}-yeast$ for 6 hours or cultured with freshwater Chlorella for 24 hours. The growth and survival rates of larval flounder fed on rotifer enriched with Marine ${\alpha}$ for 6 hours and cultured with marine Chlorella for 24 hours were higher than those of larval flounder fed on rotifer enriched with ${\omega}-yeast$ for 6 hours or cultured with freshwater Chlorella for 24 hours. And the content of n-3 HUFA of larval flounder fed on rotifer enriched with Marine ${\alpha}$ for 6 hours was higher than that of larval flounder fed on other rotifers, The results from this study indicated that rotifer culture with marine Chiorelia would be suitable for the high density culture and effective diet for the growth of larval flounder.

• ALGAE
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• v.33 no.1
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• pp.127-141
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• 2018

Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1603-1611
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• 2009

High-cell-density cultivation of yeast was investigated using the agricultural waste products corn steep liquor (CSL) and molasses. The Saccharomyces cerevisiae KV-25 cell mass was significantly dependent on the ratio between C and N sources. The concentrations of molasses and CSL in the culture medium were statistically optimized at 10.25% (v/v) and 16.87% (v/v), respectively, by response surface methodology (RSM). Batch culture in a 5-l stirred tank reactor using the optimized medium resulted in a cell mass production of 36.5 g/l. In the fed-batch culture, the feed phase was preceded by a batch phase using the optimized medium, and a very high dried-cell-mass yield of 187.63 g/l was successfully attained by feeding a mixture of 20% (v/v) molasses and 80% (v/v) CSL at a rate of 22 ml/h. In this system, the production of cell mass depended mainly on the agitation speed, the composition of the feed medium, and the glucose level in the medium, but only slightly on the aeration rate.

• Animal cells and systems
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• v.5 no.3
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• pp.205-209
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• 2001

Mesenchymal cells from the leg buds of stage 24-chick embryos differentiated into chondrocytes when plated at high density. Treatment of high density micromass culture of chick mesenchymal cells with cytochalasin D(CD, 2 $\mu$M for 24 h) resulted in inhibition of chondrogenesis. CD treatment was found to affect the expression of the contractile protein $\alpha$-smooth muscle actin ($\alpha$-SM actin). In control cultures, $\alpha$-SM actin uniformly expressed from culture day 2, but the CD-treated cells induced expression of $\alpha$-SM actin from the first day of culture followed by a continuous increase. Expression of pan-cadherin (P-cadherin) decreased as chondrogenesis proceeded in the control culture, whereas the CD-treated cells showed sustained expression. These results propose a close connection of chondrogenic differentiation with expression of $\alpha$-SM actin and P-cadherin.