• Journal of Microbiology and Biotechnology
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• 제31권3호
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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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• 제1권2호
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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.

• 한국수산과학회지
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• 제32권6호
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• pp.753-757
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• 1999

본 연구는 여러 수온 (24, 28 및 $32^{\circ}C$)에서 공기 및 산소공급과 $32^{\circ}C$에서 pH를 7로 조절된 배양에서 rotifer의 성장을 평가하였다. Rotifer는 농축 Chlorella를 먹이로 공급하였다. 또한 농축 Chlorella와 산소 및 pH를 조절한 rotifer 배양 (고밀도 배양)방법과 빵 효모와 경유 보일러를 이용한 rotifer 배양 (batch 배양) 방법간의 생산성 을 조사하였다. 공기와 산소를 공급한 배양방법에 있어서 수온이 증가할수록 rotifer의 성장률은 증가하였다. 공기를 공급한 배양에 있어서 rotifer의 최고밀도는 수온 $24^{\circ}C$에서 16,300$\~$17,000 개체/ml였고 산소를 공급한 배양에 있어서 rotifer의 최고밀도는 수온 $28^{\circ}C$에서 26,300$\~$30,500 개체/ml였다. 공기를 공급한 배양에 있어서 용존산소가 1 ppm이하로 감소하거나 산소를 공급하는 배양에서 이온화되지 않은 암모니아의 농도가 16.6$\~$22.6 ppm이상으로 증가할 때 rotifer의 성장은 감소된다. 산소공급 및 pH를 7로 조정할 때 rotifer의 최고밀도는 43,000개체/ml까지 도달한다. 고밀도 배양과 batch 배양방법에 따른 rotifer 100억개체당 생산비는 각각 693천원, 961천원이였다. 그러므로 고밀도 배양에서의 rotifer 생산성은 batch 배양한 것 보다 더 효율적이다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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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%.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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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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• 제38권4호
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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.

• 한국수산과학회지
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• 제33권2호
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• pp.93-97
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• 2000

본 연구는 먹이종류에 따라 고밀도로 배양한 rotifer를 넙치 자어에게 공급하였을 때 이들의 먹이효율을 조사하였다. Rotifer 고밀도 배양의 실험구 먹이는 18 시간 동안 담수산 Chlorella를 공급한 후 6시간 유지효모 및 $Marine\;{\alpha}$를 공급한 것과 24시간 동안 담수산 농축 Chlorella 및 해수산 농축 Chlorella를 공급한 것으로 하였다. 배양수조는 $2l$를 이용하였고 수온 $28{\circ}C$를 유지하였다. $Marine {\alpha}$와 해수산 농축 Chlorella를 공급한 실험구의 n-3 HUFA 함량이 유지효모와 담수산 농축 Chlorella를 공급한 실험구보다 높게 나타났다. 따라서 $Marine\;{\alpha}$와 해수산 농축 Chlorella를 공급한 실험구가 유지효모와 담수산 농축 Chlorella를 공급한 실험구보다 넙치 자어의 성장과 생존률이 높게 나타났다. 또한 $Marine\;{\alpha}$로 배양된 rotifer를 섭취한 넙치 자어 자체의 지방산 중 n-3 HUFA비율이 다른 먹이보다 높게 나타났지만 안정적 배양은 이루어지지 않았다. 따라서 본 실험을 통해서 rotifer 고밀도 배양시 안정적인 rotifer 성장과 rotifer의 질적 개선을 위해서는 24시간 해수산 농축 Chlorella를 공급하는 것이 효율적이며 이들을 넙치 자어에게 공급하였을 때 가장 적절한 먹이일 것으로 판단된다.

• ALGAE
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• 제33권1호
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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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• 제19권12호
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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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• 제5권3호
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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.