• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.91-95
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• 1999

To investigate the effect of salts on the production of erythritol by Torula sp., cells were grown on the media containing various concentrations of KCl or NaCl. Cell growth and glucose consumption rates decreased when KCl or NaCl concentration increased from 0.0 to 0.5M. The production of erythritol, however, was maximal at 0.3M aCl. The erythritol concentration of 54.3g/l in the medium containing 0.3M NaCl and 200g/l glucose was obtained after 120h. The production of erythritol decreased in cultures above 0.3M NaCl or 0.4M KCl due to the inhibition of cell growth. To elucidate the effect of salts more quantitatively, KCl and NaCl concentrations were converted to osmotic pressure. As the osmotic pressure increased, the yield of erythritol from glucose increased regardless of the kinds of salts and the yield of erythritol was approximately 49% at the osmolality of 2.4Os/kg. When the osmotic pressure increased to 2.5Os/kg, the specific growth rate of cells decreased but the production rate of erythritol increased. For the effective production of erythritol, osmotic pressure must be adjusted not to inhibit markedly the growth rate of cells and to stimulate the production rate of erythritol by supplementing salt.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.374-378
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• 2004

The effects of various environmental factors such as pH (5, 6, 7, 8 and 9), temperature (30, 37 and $40^{\circ}C$) and rotational speed (150, 200 and 250 rpm) on the growth and the hepatitis B core antigen (HBcAg) production of Escherichia coli W3110IQ were examined in the present Study. The highest growth rate is achieved at pH 7, $37^{\circ}C$ and at a rotational Speed of 250 rpm which is 0.927 $h^{-1}$. The effect of pH on cell growth is more substantial compared to other parameters; it recorded a $123\%$ different between the highest growth rate (0.927 $h^{-1}$) at pH 7 and lowest growth at pH 5. The highest protein yield is achieved at pH 9, rotational speed of 250 rpm and $40^{\circ}C$. The yield of protein at pH 7 is $154\%$ higher compared to the lowest yield achieved at pH 5. There is about $28\%$ different of the protein yield for the E. coli cultivated at 250 rpm compared to that at 150 rpm which has the lowest HBcAg yield. The yield of protein at $40^{\circ}C$ is $38\%$ higher compared to the lowest yield achieved at $30^{\circ}C$.

• 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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.142-146
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• 2003

Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

• KSBB Journal
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• v.11 no.2
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• pp.254-261
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• 1996

Attenuated varicella-zoster virus (VZV) was propagated in human lung fibroblast (HLF) cells Among media tested in this work, DMEM was the best medium for the growth of HLF cells. Because HLF was a normal finite cell line, cell growth late was dependent on the age of HLF cells. When the population doubling level (PDL) was higher than 46, apoptosis of HLF cells started and cell growth rate decreased. The optimum temperature for the cell growth and virus propagation in the T-flask culture was $37^{\circ}C$. In a microcarrier culture system in which Cytodex-3 was used for the VZV propagation in spinner vessels, the yield of plaque forming cells was lower than that in the T-flask culture. The relatively high shear environment near microcarriers was thought to cause the low yield of VZV in the microcarrier culture system.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Archives of Pharmacal Research
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• v.28 no.2
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• pp.220-226
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• 2005

Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcar-bamoylase (OTC). When supplements were added to the medium at $33\;^{\circ}C$, the growth of IBE and CO5 cells increased by $27\%\;and;26\%$, respectively and the maximum yield of EPO was increased by $40\%$ in both cell lines. The absolute EPO concentration in the CO5 cells was always $55{\sim}60\%$ higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at $33\;^{\circ}C$ until their growth rates approached those at $37\;^{\circ}C$, the growth rates of both IBE and CO5 cells increased by $54\%$ and their maximum EPO levels increased by up to $73\%\;and\;56\%$, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.

• KSBB Journal
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• v.13 no.6
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• pp.644-648
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• 1998

In the production of a low cost bacterial insecticide, it is important to produce a high spore concentration using low price substrates. Experiments were carried out to investigate the effects of the addition of mineral salts and glucose, and of dissolved oxygen concentration on the cell growth and spore formation of Bacillus thuringiensis var aizawai using a cheap wheat and soybean meal in the batch culture. The maximum viable cell number was 1.2${\times}$109 CFU/mL at 12 hr culture and spore yield was 54.2% at 74 hr culture using an industrial medium containing 20 g/L wheat meal and 30 g/L soybean meal under 1.0 vvm aeration and 200 rpm agitation. The cell growth and the spore formation were not enhanced by the addition of mineral salts in industrial medium, whereas th addition of 10g/L glucose decreased the cell growth and spore formation. We could obtain a maximum viable cell number of 2.2${\times}$109 CFU/mL and spore number of 1.9${\times}$109 CFU/mL at the dissolved oxygen concentration of 60% of saturation. The spore concentration was enhanced approximately by 2 times as compared to the dissolved oxygen concentration of 50%. In the bench-scale culture, the maximum viable cell and spore number were 2.5${\times}$109 CFU/mL, and 2.2${\times}$109 CFU/mL, respectively under 1.0 vvm aeration and 400 rpm agitation. The spore yield was 88% based on the maximum viable cell number. As a result, it was confirmed that the production of high spore concentration could be obtained by a bench-scale culture using an industrial medium.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.173-177
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• 1989

Cell density was gradually decreased as the dilution rate was increased under chemostat cultivation of HeLa cells. Maxium cell density was maintained at the dilution rate of 0.020 (1/h) which was far less than the wash-out rate of 0.050 (1/h), Maxium cell productivity of 2 (mL of cells/L/h)was obtained at the dilution rate of 0.030 (1/h) by showing the culture also required maintenance period at low dilution rates, whose result meant the deviation of continuous culture theroy. Methods of indirectly measuring cell density have been introduced to represent mammalian cell growth, which are packed cell volume and oxygen uptake rate, and these values showed good linear relationship with actual cell density by having correlation factor of 0.90. Theoretical maximum oxygen yield, $Y_{O2}^{max}$ and maintenance oxygen consumption rate, m$_{O2}$, were estimated as 4.1$\times$10$^5$ (cells/mmole $O_2$) and 10.71$\times$10$^{-9}$ (mmole $O_2$/cells/h) by employing oxygen yield model.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1677-1680
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• 2010

The harmful effects of succinic acid and oxidative stress on cell growth were determined during batch fermentation with Mannheimia succiniciproducens LPK7, a powerful succinic acid-producing strain, and conditions were optimized to minimize these effects. In terms of toxicity, the cell concentration decreased as the concentration of succinic acid increased. By changing the pH from 6.5 to 7 during fermentation, the cell concentration increased by about 10%, and the level of succinic acid production was 6% higher than that of the control. In addition, by introducing protectants, the cell concentration increased by about 10%, and the level of succinic acid produced was increased by 3%.