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Kinetic Analysis of the Effect of Cell Density on Hybridoma Cell Growth in Batch Culture

  • Lee, Eun-Yeol
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.117-120
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    • 2002
  • The effect of cell density on cell growth was investigated in a suspension batch culture of hybridoma cells. The specific growth rate was found to increase with increasing initial cell density and then to decrease with further increases in initial cell density. In order to quantitatively describe the dependence of specific growth rate on cell density, a kinetic model is proposed, which satisfactorily represents the experimental data.

Application of Computer-coupled Mass Spectrometer for Continuous On-line Monitoring of Cell Growth and Growth Rate (세포증식과 증식속도의 On-line Monitoring을 위한 Computer- coupled Mass Spectrometer의 응용)

  • 남수완;최춘순;김정회
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.241-246
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    • 1989
  • Continuous on-line monitoring of cell concentration and growth rate in aerobic batch fermentation process was carried out by analyzing the exhaust gas composition of tormentor with a quadrupole mass spectrometer. From the mass spectrometric analyses of major gaseous components, i.e. $N_2$, $O_2$, $CO_2$ and $H_2O$, and the material balance equations for oxygen and carbon dioxide, oxygen uptake rate (OUR) rind carbon dioxide evolution rate (CER) were instantaneously calculated using a computer (16-bit IBM PC-AT) interfaced to a quadrupole mass spectrometer. The calculated OUR and CER data were used for the estimation of cell concentration and growth rate of Candida utilis during batch culture. It was found that the cell concentration could be satisfactorily estimated from the data of OUR arid CER during the culture and this method could be successfully und for the continuous monitoring of cell growth and growth rate.

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BcI-2 Over-expression Reduced the Serum Dependency and Improved the Nutrient Metabolism in a NS0 Cells Culture

  • Tey Beng Ti;Al-Rubeai Mohamed
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.254-261
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    • 2005
  • The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The ${\mu}_{max}$ and $K_s$ for the Bcl-2 cell line is $0.927day^{-1}\;and\;0.947\%(v/v)$ respectively, which are $21\%$ greate and $7\%$ lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a $17\%$ decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EM suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.

Effect of Incorrectly Estimated Parameters on the Control of Specific Growth Rate in E. coli Fed-Batch Fermentation

  • Park, Tai-Hyun;Yoon, Sung-Kwan;Kang, Whan-Koo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.22-25
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    • 1996
  • An Exponetial feeding strategy has been frequently used in fed-batch fermentation of recombinant E. coli. In this feeding scheme, growth yield and initial cell concentration, which can be erroneously determined, are needed to calculate the feed rate for controlling specific growth rate at the set point. The effect of the incorrect growth yield and initial cell concentration on the control of the specific growth rate was theoretically analyzed. Insignificance of the correctness of those parameters for the control of the specific growth rate was shown theoretically and experimentally.

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Mathematical Analysis of a High Density Animal Cell Culture with a Spin-Filter (회전식 여과기를 이용한 고농도 동물세포배양의 수학적 해석)

  • 박흥우
    • KSBB Journal
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    • v.9 no.2
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    • pp.230-237
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    • 1994
  • Spin-filters are used as cell separation devices for achieving high cell density and high productivity in animal cell culture. We have proposed a model for the cell growth in a spin-filter perfusion culture and examined the effects on cell growth by several parameters including ammonia inhibition, specific growth rate, specific feeding rate, and cell retention. Results from computer simulation and sensitivity analysis indicate that the cell retention affects the cell growth mostly while there is a significant inhibition on cell growth by the ammonia accumulated during the culture. The specific feeding rate has minimal effects on cell growth, which is consistant with the fact that the cell growth with a step feeding is quite similar to that with a continuous feeding.

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High Cell Density Culture of Anabaena variabilis with Controlled Light Intensity and Nutrient Supply

  • Yoon, Jong-Hyun;Shin, Jong-Hwan;Ahn, Eun-Kyung;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.918-925
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    • 2008
  • Controlling the light energy and major nutrients is important for high cell density culture of cyanobacterial cells. The growth phase of Anabaena variabilis can be divided into an exponential growth phase and a deceleration phase. In this study, the cell growth in the deceleration phase showed a linear growth pattern. Both the period of the exponential growth phase and the average cell growth rate in the deceleration phase increased by controlling the light intensity. To control the light intensity, the specific irradiation rate was maintained above $10\;{\mu}mol/s/g$ dry cell by increasing the incident light intensity stepwise. The final cell density increased by controlling the nutrient supply. For the control of the nutrient supply, nitrate, phosphate, and sulfate were intermittently added based on the growth yield, along with the combined control of light intensity and nutrient concentration. Under these control conditions, both final cell concentration and cell productivity increased, to 8.2 g/l and 1.9 g/l/day, respectively.

Modeling of Typical Microbial Cell Growth in Batch Culture

  • Jianqiang Lin;Lee, Sang-Mok;Lee, Ho-Joon;Koo, Yoon-Mo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.382-385
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    • 2000
  • A mathematical model was developed, based on the time dependent changes of the specific growth rate, for prediction of the typical microbial cell growth in batch cultures. This model could predict both the lag growth phase and the stationary growth phase of batch cultures, and it was tested with the batch growth of Trichoderma reesei and Lactobacillus delbrueckii.

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Measurement of Cell Death Constant in Anabaena flos-aquae (Cyanophyceae) by the Molecular Probe (Anabaena flos-aquae 에서의 세포사멸계수(Cell Death Constant)의 측정)

  • 오인혜
    • The Korean Journal of Ecology
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    • v.20 no.3
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    • pp.169-173
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    • 1997
  • The measurement of cell death constant in Anabaena flos-aquae was tested by the Live/Dead BacLight Viability kit(Molecular Probes Co., Seatle, WA). When the Live/Dead BacLight Viability kit was applied to Anabaena flos-aquae, the cells with intact cell membranes(live cells) stained fluorescent green, while the cell with damaged membranes(dead cells) stained fluorescent red and the background remained virtually nonfluorescent. The rations of live : dead cells in the cell suspension were controlled artifically and Live/Dead BacLight Viability kit was applied to them. The ratios of green:red fluorescent cells in the cell suspension were the same as those of live : dead cells controlled artifically. It was also approved by the fluorescence emission. The cell death constant was measured in the P-limited Anabaena flos-aquae chemostal culture in the N-fixing and $KNO_3-supplied$ conditions. The culture in N-fixing chemostat had a dead cell proportion of 1.2% at the growth rate of 0.7/day and increased to 2.6% at the growth rate of 0.3/day. The cell death constant of N-fixing culture was 0.008/day.There was a same trend in the $KNO_3-supplied$ chemostat culture. The proportion of dead cell was 1.6% of dead cell proportion at the growth rate of 0.7/day and increased to 4.3% at the growth rate of 0.3/day.

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Effects of Plug Cell Trays, Soil and Shading Rates on Seed Germination and Seedling Growth Characteristics of Hippophae rhamnoides L.

  • Lee, Songhee;Cho, Wonwoo;Chandra, Romika;Han, Jiwon;Kang, Hoduck
    • Journal of Forest and Environmental Science
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    • v.36 no.1
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    • pp.55-61
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    • 2020
  • In this study, basic data with respect to the introduction of Hipphophae rhamnoides L. and its cultivation in Korea could be obtained. According to the size of the plug cell tray, Chinese origin's rate of seed germination was relatively high in 128 plug cell tray, and growth was vibrant in 50 plug cell tray. The germination and growth of Russian origin seeds showed that they were relatively effective in 50 plug cell tray and with respect to soil environment, TKS-2 soil with untreated shading relatively promoted both germination and growth for Chinese origin, the rate of germination was high in bed soil for horticulture and growth result was good in TKS-2 in the case of Russian origin. It was confirmed that the germination rate of Chinese origin H. rahmnoides L. was highest in untreated shading and the shoot growth was vibrant in 70% shading while the growth in roots was vibrant in the untreated shading. In the Russian origin, H. rhamnoides L. the germination rate in 30% and 70% shading was about 50% which was higher than that in the untreated shading and general growth was vibrant in 30% shading.

Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.110-117
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    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

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