• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.51-54
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• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• Journal of Microbiology and Biotechnology
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• 제16권5호
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• pp.673-677
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• 2006

A perfusion culture of transgenic Nicotiana tabacum cell suspensions, transformed to express recombinant glucuronidase (GUS), was successfully performed in a 5-1 stirred tank bioreactor. With 0.1 $day^{-1}$ of perfusion rate, the maximum dry cell weight (DCW) reached to 29.5 g/l in 16 days, which was 2.1-fold higher than the obtained in batch culture (14.3 g/l). In terms of the production of GUS, the volumetric activity could be increased up to 12.8 U/ml by using perfusion, compared with 4.9 U/ml in batch culture. The specific GUS activities in both perfusion and batch cultures were maintained at similar levels, 200-400 U/g DCW. Consequently, a perfusion culture could be a good strategy for the enhanced production of recombinant proteins in a plant cell culture system.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.323-327
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• 1999

It was shown that brain-derived neurotrophic factors (BDNFs) secreted from human neuroblastoma cells can significantly improve the growth of the neurites of PC12 nerve cells. The addition of purified BDNFs elongated the neurites of PC 12 nerve cells two to three times more than the case where the addition was not made. The perfusion rate strongly affected the change of the size of human neuroblastoma cells because the cell size decreased as the perfusion rate increased. This could also influence the productivity of BDNF from the cells. It is also important to note that the BDNF production was decreased when the cell size was reduced. BDNF production rate also decreased at a fast perfusion rate in a smaller cell size. At the relatively fast perfusion rate of 18 ml/h, the ratio of apoptotic to necrotic cells dramatically decreased, which possibly caused the decrease of BDNF production. It has been proven that the secretion of BDNF from human neuroblastoma cells was a partially growth-related process by yielding 6.2$\times l0^{-8}/g$ of BDNF/cell/h of growth related parameter and $0.48{\times}l0^{-9}/g$ of BDNF/cell/h of nongrowth-related parameter in a growth kinetic model. In addition, it was also found that the perfusion rate played a very important role in controlling the cell death mechanism.

• 한국미생물·생명공학회지
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• 제18권1호
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• pp.61-65
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• 1990

Murine hybridoma 세포배양시 lectin 계통의 생산 증진제가 무혈청 배지의 단계적 유입에 의해 약 40mg/l의 단일항체를 생산함으로서 단일항체의 생산성을 증가시킬 수 있음이 확인됐으며, 또한 이 배양공법으로 무혈청 배지에서 연속배양이 불가능한 세포의 배양이 가능했다. 이같은 무혈청 배지의 단계적 유입공법으로 약 28 x $10^{-10}$mg/cells/h의 속도로 단일항체가 생산된 것에 비해, 10 혈청이 포함된 배지로는 연속 배양시 11.1 x $10^{-10}$mg/cells/h 의 속도로 생산됐으며 회분배양의 경우는 4.0 x $10^{-11}$mg/cells/h의 속도로 생산됐다.

• KSBB Journal
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• 제4권1호
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• pp.34-39
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• 1989

본 논문은 동물 세포의 대량 배양을 위해 연속 공법 방식인 Perfusion Continuous System을 도입해 의약적으로 중요한 EPO의 생산을 위한 생물 공학적인 자료들을 제고하고 있다. 이 System은 세포 증식 속도를 배지의 Perfusion 속도로 변화시킴으로써 조절시킬 수 있는 산소 소비속도와 밀접한 상관관계가 있음을 입증하며 이는 세포수의 직접 측정에 따른 오차 및 방법상의 문제를 정학히 측정할 수 있는 간접 변수, 즉 산소소비속도를 이용함으로써 제거할 수 있다. 특히 이 산소소비속도와 세포 성장 관계로 model로써 세포 증식을 예측함과 동시에 동물 세포 대량 배양을 위한 scale-up의 중요한 기초자료가 될 것이다. 지금까지 발표되지 않았던 동물 세포의 glucose에 대한 True growth yield와 maintenance coefficient값들의 측정은 동물 세포 성자관 유용 물질 생산을 위한 중요한 수율적 자료가 된다.한편 이 결과는 지금까지 미생물이나 광합성에서만 적용되었던 yield model이 Eukaryotes에서도 응용될 수 있음을 증명하고 있다. 이와 같이 perfusion system이 많은 장점을 갖고 있지만, 세포 성장에 따른 동력학적인 연구의 수행이 좀 더 요구되는 실정이며, 특히 Perfusion system을 설명할 수 있는 이론 및 cytostatic moel의 정립이 선행되어야 할 것이다.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.457-462
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• 2002

Low Electromagnetic Field (EMF) intensity in the range of $1{\mu}T\;to\;10{\mu}T$(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At $1{\mu}T\;intensity,\;1.3{\times}10^7$ viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to $2.8{\times}10^6$ viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain $1.2{\times}10^7$ viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between $1{\mu}T\;and\;10{\mu}T$ in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at $1{\mu}T$EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of $Ca^2+$ was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than $10{\mu}T$ could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.401-405
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• 1990

5 혈청을 포함한 DMEM 배지에 갑상세포주 579를 연속배양하여 약 $5.7 \times 10^{-8}$g/h /cell에 해당하는 pro-UK의 비생산속도를 얻을 수 있었다. 또한 배지의 이동속도가 증가할 수록 glutamine의 소비속도가 증가하는 반면 ammonia 생산속도는 일정하게 유지되는, glutamine의 완전동화에의한 물질생산증가 현상을 나타냈다. 5mM의 glucose와 2mM의 glutamine, 포화용존 공기의 10에 해당하는 용존산소 및 pH 6.2의 maintenance 생육 조건하에서 약 15일간 유지시켜, $12\times 10^{-8}$g of pro-UK/h/cell의 최대 비생산속도와 0.226mg/g of glucose의 생산수율을 얻었으며, 이는 10ml/min의 배지 이동속도를 유지하는 연속배양 조건에서 매일 0.223mg의 pro-KU를 생산할 수 있을을 의미한다.

• KSBB Journal
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• 제8권1호
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• pp.17-22
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• 1993

$1.96{\times}10^{-5}$(IU/cell/hr) of specific scu-PA production rate was obtained from HEK cells in maintaining ca. $8{\times}10^{5}$(cells/ml) of maximum roll density at 10(ml/hr) of perfusion rate with recycling 20% serum free conditioned media. It can be compared to $4{\times}10^{6}$(cells/ml) of maximum cell density and $4.56{\times}10^{-4}$(IU/cell/hr) of specific production rate in cultivating cells with 1% serum containing medium. Thc conversion ratio of scu-PA to tc-UK increased up to 55% as the recycling ratio increased; however, recycling the used medium seemed to have least negative effect on cell growth. It also showed that the recycling process had definitive advantage of using serum free medium in perfusion cultivation of HEK cell line.

• 한국미생물·생명공학회지
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• 제19권5호
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• pp.504-508
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• 1991

유전자 재조합된 동물 세포의 유가 배양시 $1.85\times 10^{-10}$(mmole/cell/h)의 비 glucose 소비속도와 $4.7\times 10^{-7}(\mu g/ceil/h)$의 erythropoetin (EPO)비 생산속도를 유지할 수 있었다. 또한 이같은 배양에서 회분 및 연속배양에서 보다 높은 세포수를 얻었으며 전 배양이 유사 안정상태에 도달하는 배양 후기에는 glutamolysis가 생육 공정에 매우 중요한 역활을 하고 있음이 확이됐다. 유가 배양시 13(mmloe/l)의 glucose 농도에서 생육 제한 현상이 일어났으며, 이같은 농도에 도달할 때까지는 glucose의 농도가 증가함에 따라 배양시간의 경과와 함께 EPO 생산성이 증가했다.

• KSBB Journal
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• 제21권5호
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• pp.336-340
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• 2006

본 연구에서는 참당귀 현탁세포배양에 의한 면역 증강성 ECP의 생산을 증진시키기 위하여 배지교환식 배양을 수행하였으며, ECP의 분비를 촉진하는 초음파 처리와 Pluronic F-68의 영향을 조사하였다. 참당귀의 현탁세포배양시, 최대 세포생장은 6일째에 16.8 g DCW/L였고, ECP의 생산은 세포생장과 함께 증가하다가 8일째에 최고 0.9 g/L가 생산되었다. 식물세포 고농도배양법인 배지교환식 배양을 적용해, 초기 당 농토를 높이고 접종 후 5일부터 연속적으로 배지교환을 해주어 23.8 gDCW/L의 고농도의 현탁세포를 얻을 수 있었다. 초음파 처리 및 Pluronic F-68의 첨가는 참당귀 세포의 세포막의 투과성을 증진시켜 ECP의 생산성을 높임과 동시에 세포 내의 다당의 배지로의 배출을 유도한 것으로 판단된다.