• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.189-194
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• 2009

The objective of this study was to determine effects of different culture media. Preantral follicles were mechanically extracted from bovine ovaries and cultured for 16 days in tissue culture medium (TCM)-199, DMEM or alpha-minimal essential medium ($\alpha$-MEM) + 10% FBS + 0.1 mg/ml sodium pyruvate + 100 mIU/ml FSH. The collected primary follicles from ovary were higher than the primary and secondary follicles. The survival rates of the follicles in TCM-199 were significantly higher (p<0.05) than those in DMEM and $\alpha$-MEM. The diameter of the follicles progressively increased during 12 days of culture. The maximum size ($139.1{\pm}5.4\;{\mu}m$) reached on Day 12 of the in vitro culture and decreased on Day 16. These results suggest that in a culture of bovine preantral follicles, TCM-199 is an optimal medium and a longer-term culture of preantral follicles (>12 days) may be needed to form antra.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.1
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• pp.86-93
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• 2020

The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

• Journal of Bio-Environment Control
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• v.14 no.4
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• pp.269-274
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• 2005

The purpose of this experiment was to investigate optimal ionic concentration of nutrient solution for water culture of young welsh onion (Allium fistulosum). For the purpose of clarification of optimal nutrient concentration to maximize growth of young welsh onion, different nutrient concentrations of Yamazaki's solution for welsh onion seedling $(NO_3^--N\;9.0,\;NH_4^+-N\;3.0,\;PO_4^{3-}-P\;6.0,\;K^+7.0,\;Ca^{2+}\;2.0,\;Mg^{2+}\;2.0,\;and\;SO_4^{2-}-S\;4.4me{\cdot}L^{-1})$ which selected by prior experiment were treated as 0.6, 1.2, 1.8, and $2.4dS{\cdot}m^{-1}$. Increments of fresh weight, dry weight and top length were the highest in 1.2 and, in the next, were placed by the order of 1.8, 2.4, and $0.6dS{\cdot}m^{-1}$ The regression coefficients for the maximal growth of fresh weight of cv. 'Geurnjanguedaepa' and 'Tokyokuro' were $y=-42.091x^2+171.79x+11.047 (R^2=0.8946,\; R=0.9458^*)\;and\;y=-50.069x2+157.58x+15.414(R^2=0.9343,\;R=0.9692^{**})$, respectively, and optimal EC levels according to regression coefficients were 1.68 and $1.57dS{\cdot}m^{-1}$. As the conclusions, optimal nutrient levels far young welsh onion were $1.2dS{\cdot}m^{-1}$ EC in the early growth stage and $1.6\~l .7dS{\cdot}m^{-1}$ in the later growth stage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.3
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• pp.373-378
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• 2008

The objective of this paper was to investigate optimal culture conditions for the production of an inhibitor against subtilisin-like protease from Streptomyces thermocarboxydus (S. thermocarboxydus) C12 isolated from sediments of Gwangyang coast. The optimal temperature and initial pH for the production of subtilisin-like protease inhibitor were $40^{\circ}C$ and pH 8.0, respectively. Inhibition activities were high for galactose, glucose and fructose. The best carbon source and its concentration were galactose and 1.6% (w/v), respectively. Inhibition activities were also high in medium containing polypeptone, proteose and peptone. Optimal nitrogen source and concentration were protease peptone and 0.5% (w/v), respectively. Optimal concentrations for inhibitor production were 1% (w/v) for NaCl and 1 mM LiCl for metal salts. The subtilisin-like protease inhibitor from S. thermocarboxydus C12 showed a maximum inhibitor activity after cultivation for 84 h under the optimized medium.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.5
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• pp.275-280
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• 2002

In order to control glucose concentration during fed-batch culture for antibiotic production, we applied so called “software sensor” which estimates unmeasured variable of interest from measured process variables using software. All data for analysis were collected from industrial scale cultures in a pharmaceutical company. First, we constructed an estimation model for glucose feed rate to keep glucose concentration at target value. In actual fed-batch culture, glucose concentration was kept at relatively high and measured once a day, and the glucose feed rate until the next measurement time was determined by an expert worker based on the actual consumption rate. Fuzzy neural network (FNN) was applied to construct the estimation model. From the simulation results using this model, the average error for glucose concentration was 0.88 g/L. The FNN model was also applied for a special culture to keep glucose concentration at low level. Selecting the optimal input variables, it was possible to simulate the culture with a low glucose concentration from the data sets of relatively high glucose concentration. Next, a simulation model to estimate time course of glucose concentration during one day was constructed using the on-line measurable process variables, since glucose concentration was only measured off-line once a day. Here, the recursive fuzzy neural network (RFNN) was applied for the simulation model. As the result of the simulation, average error of RFNN model was 0.91 g/L and this model was found to be useful to supervise the fed-batch culture.

• Journal of Forest and Environmental Science
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• v.24 no.1
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• pp.53-59
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• 2008

Extracellular enzyme activities of Fomitopsis palustris were determined by the particle sizes, culture periods and concentrations of wood particle substrate which was mixture of 4 domestic coniferous woods, such as Pinus densiflora, Larix leptolepsis, Pinus koraiensis, and Pinus rigida. The results showed that the culture conditions had an effect on the secretion of most of the extracellular enzymes from Fomitopsis palustris in the mixed wood particle substrate. :The optimal culture conditions for enzyme activities were 80~100 mesh in wood particle size, 7.5% in concentrations of wood substrate, and 4~8 weeks in culture period.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.310-318
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• 2012

We conducted a screening and checked the cultivation methods of lactic acid bacteria, which have anti-atopic dermatitis functions, by determining the lactic acid bacteria's immune enhancement by FACS, and antimicrobial activity against Staphylococcus aureus. The increase of Tcell CD4+/CD25+/foxp3+ was bigger in Lactobacillus plantarum than Lactococcus lactis subsp. lactis (Lc. lactis) and the antimicrobacterial activity against S. aureus was the opposite. The antimicrobial activity of Lb. plantarum culture with medium containing Lc. lactis culture broth was not enhanced, but the antimicrobial activity of Lc. lactis cultured in a medium containing Lb. plantarum culture broth was enhanced. As the optimal method caltivation of Lc. lactis in a medium containing 10% of heat-killed Lb. plantarum culture broth was chosen. By this method, the antibacterial activity of the pure Lc. lactis culture increased sharply at the end of the log phase, while a restraint effect on the growth of S. aureus increased 1.29 times.

• KSBB Journal
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• v.25 no.2
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• pp.187-192
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• 2010

We designed the optimal operational strategy in repeated fed-batch ethanol fermentation using Sacchromyces cerevisiae ATCC 24858 in views of ethanol yield, specific ethanol production rate, and ethanol productivity, when the aeration rate were controlled at 0.0 and 0.33 vvm. Coincidentally, the time intervals of withdrawal-fill of culture medium (24 and 36 h) were investigated. Ethanol yield and ethanol productivity when the aeration was carried out at 0.33 vvm were superior to those when the aeration was not carried out. Additionally, those parameters when the time interval of withdrawal-fill of culture medium was 24 h were superior to those when time interval of withdrawal-fill of culture medium was 36 h. The total ethanol production reached at the greatest value, 703.8 g-ethanol, when the aeration was carried out at 0.33 vvm and the time interval of withdrawal-fill of culture medium was 24 h. In this study, we verified experimentally the necessity of designing the operational strategy for increasing ethanol production in terms of aeration rate and time interval of withdrawal-fill of culture medium in the repeated fed-batch ethanol fermentation.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.37-40
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• 1999

This study was carried out to determine the optimal conditions for the production of plants derived from the unpollinated ovary culture of Chinese chive (Allium tuberosum Rottl.). The Chinese chive collected from Korea showed much higher frequency of plantlet formation than those from Japan in the culture of unpollinated ovaries. Among the collections, 'Youngiljaerae' showed the highest frequency of plantlet formation. The MS basal medium was superior to B/sub 5/ in plantlet formation. The ovaries inoculated on the 2,4-D-free medium were directly induced plantlets without callus formation. Floral parts inoculated as a unit played important roles in callus formation and plant production. The frequency of callus and plantlet formation was higher in the culture of ovary with anthers than that of ovary alone.