• 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.

• International journal of thyroidology
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• 제11권2호
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• pp.160-166
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• 2018

Background and objectives: Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system. Materials and Methods: We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish. Results: Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish. Conclusion: This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

• 대한기계학회논문집B
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• 제35권11호
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• pp.1127-1131
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• 2011

본 논문에서는 액적의 균형공급에 의해 관류유량이 일정하게 유지되는 펌프 없는 세포배양 칩을 제안하였다. 기존의 펌프 없는 세포배양 칩은 유체 수위차가 시간에 따라 점차 감소하여 일정한 관류유량 유지가 어려웠다. 반면, 제안된 칩은 액적의 균형공급으로 유체 수위차를 일정하게 유지하여 일정한 관류유량의 세포배양이 가능하다. 제작된 세포배양 칩의 성능분석 결과, 펌프 없이 최대 9.96%와 6.92% 의 편차 및 오차 내에서 0.1~$0.3{\mu}l$/min 의 관류유량, Q, 을 얻었다. H358 폐암 세포주 배양결과, Q=$0.2{\mu}l$/min 의 관류유량에서 최대 $57.8{\pm}21.1%$/일의 증식률을 보여, Q=$0{\mu}l$/min 의 정치배양보다 1.9 배 높은 값을 얻었으며 활성도 또한 정치배양보다 관류배양이 더 높은 값을 보였다. 제안된 펌프 없는 세포배양칩은 높은 증식률과 활성도의 좋은 배양환경을 제공하여 세포기반 바이오 분석에 응용 가능하다.

• KSBB Journal
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• 제13권4호
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• pp.411-417
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• 1998

The possibility of application of membrane type immobilized adsorbent to the fed-batch or perfusion culture system with anchorage-independent cells as well as batch system was investigated. The improvement in cell density and cell viability due to the combination of immobilized adsorbent with each culture system was evaluated for the investigation, and the optimum culture system employing immobilized adsorbent system was suggested based on the results. It was observed that the system with immobilized adsorbent showed better cell growth and cell viability than that without immobilized adsorbent in every operation system of batch, fed-batch, and perfusion. In case of batch system, 200% improvement of maximum cell density was observed in the system where ammonium chloride was added on purpose. And 50% improvement of maximum cell density was observed in the fed-batch system where ammonium ion accumulates significantly, while small increase in maximum cell density was observed in the perfusion system where dilution of waste byproducts exists. Especially, the fed-batch system showed the most significant improvement on cell growth because both compensation of nutrient and removal of ammonium ion occurred simultaneously in the system. Therefore a combined system of immobilized adsorbent and fed-batch operation could be suggested as an optimum system with in situ removal of ammonium ion.

• KSBB Journal
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• 제14권6호
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• pp.665-671
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• 1999

본 연구에서는 식물 유래의 항암제인 taxol을 생산하는 주목세포의 현탁배양을 통하여 taxol 반합성의 전구체인 10-deacetylbaccatin III(10-DAB)의 생산을 증대시키고자 하였다. 이 차대사산물의 생산이 세포생장과 관련이 있으므로 세포생장이 우수한 배지를 선택하기 위한 실험을 수행하였고, SH 배지에서 가장 우수한 세포생장이 관찰되었다. 또한 선택된 SH 배지에서 우수한 세포생장과 10-DAB 생산을 얻기 위해 초기 접종농도와 당농도의 변화를 관찰하였다. 초기 접종농도와 당농도가 낮을수록 세포생장이 빠르며, 낮은 초기 당농도와 초기 접종농도 6 g/L FCW 이었을 때 10-DAB의 생산이 우수하였다. 생산성을 높이기 위한 고농도 세포생장과 배지로의 10-DAB 분비를 목적으로 perfusion culture를 도입하였다. 고농도 당을 첨가한 배지로 perfusion을 수행한 결과 배양 후반까지 세포생장이 가능하였고, FCW/DCW ratio가 감소되어 대조구의 2.5배인 34.67 g/L의 높은 세포농도를 얻을 수 있었다. 10-DAB의 생산은 perfusion 시작후 10일 동안은 배지로 배출된 양이 세포내보다 높았다.

• 한국정밀공학회지
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• 제28권4호
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• pp.526-533
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• 2011

This research is about the pulsatile pump system utilized in the perfusion bioreactor for the in vitro human tissue culture. A pulsatile pump system which can be applied to the culture of the vascular tissues including blood vessel is developed by using the idea of human heart's blood pumping into organs as followings: culture chamber, a pressurizing device which generates laminar pulsatile flow by controlling the x-sectional area of the culture media delivering tubing, a compliance chamber which supplies the pressuring device with a constant pressure, and a peristaltic pump which circulates the culture media in a circuit ranging from the culture chamber to the compliance chamber. The developed pulsatile pump system shows that a physiology of the human heart's blood pumping including pulsatile pressure waveform of systolic-diastolic pressure is well represented. Not only time domain but also frequency domain characteristics of pulsatile pump system which are necessary for the vascular tissue culture such as pulsatile pressure waveform's shape, the frequency, and the magnitude can be easily generated and manipulated by using the proposed system.

• KSBB Journal
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• 제13권3호
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• pp.259-267
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• 1998

Optimal feeding strategies in bioreactor operation of Scutellaria baicalensis G. plant cell culture were investigated to maximize the production of flavone glycosides by using a structured kinetic model which can predict culture growth and flavone glycosides synthesis in a rigorous, quantitative manner. For the production of baicalin and wogonin-7-0-GA, the strategies for glucose feeding into Scutellaria baicalensis G. plant cell culture were proposed based on the model, which are a periodic fed-batch operation with maintenance of cell viability and of specific production rate respectively, and a perfusion operation with maintenance of specific production rate for baicalin and wogonin-7-0-GA. Simulation results showed that the highest volumetric concentration of flavone glycosides was obtained in a periodic fed-batch operation with maintenance of cell viability among all the suggested strategies. In the periodic fed-batch operations, the higher volumetric production of flavone glycosides was achieved compared with that in the perfusion operation. It can be concluded that a periodic fed-batch operation with maintenance of cell viability would be the optimal and practical operating strategy of Scutellaris baicalensis G. plant cell culture for the production of flavone glycosides.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.130-135
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• 2000

The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1${\times}$107 cells/mL. When the perfusion rate was increased, the intermittrnt bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific oxygen consumption rate of intermittent bleeding culture. finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15L bioreactor after one-month operation.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.301-304
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• 2002

참당귀 (Angelica gigas Nakai) 현탁세포의 고농도 배양을 위하여 perfusion 배양을 한 결과 세포의 생장이 증진됨을 확인하였다 . 배양 5 일째부터 sucrose를 4배 농축한 배지로 2 mL 씩 교환해준 결과, 세포의 생장이 계속 유지되었고 최대 세포농도도 23.7 g/L 로 대조구에 비하여 1.7 배 증가하였다. 초기 당농도를 50 g/L 로 하여 배지 교환을 한 결과, 최대 세포농도도 23.8 g/L 로 30 g/L 당농도 대조구에 비하여 1.6 배 증가하였고, 배지교환을 하지 않은 50 g/L 대조구에 비하여 1.1 배 증가하였다 . 따라서 초기 당농도를 높이고 연속적으로 배지교환을 해준다면, 고농도의 세포를 얻을 수 있으며 결과적으로 참당귀 세포가 생산하는 다당류의 생산량을 증진시킬 수 있을 것이다.