• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.551-563
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• 1996

This study investigated the properties of primary cultured proximal tubule cells in hormonally defined(insulin, transferrin, and hydrocortisone), serum-free medium or 10% serum-supplemented medium. The growth rate of the primary cultured proximal tubule cells was lower in the hormonally defined, serum-free medium than in the 10% serum- supplemented medium(p < 0.05), while the activities of brush border marker enzymes, alkaline phosphatase(AP), leucine aminopeptidase(LAP), and y-glutamyl transpeptidase(${\gamma}$-GTP) were increased(p < 0.05). The activities of these enzymes, however, decreased with the lapse of incubation time to 50-70% after 6 days culture compared to those of the freshly-prepared proximal tubules. The enzymatic activities of the primary cultured proximal tubul cells on 6, 9, 12, and 15 days of culture were significantly increased in the hormonally defined, serum-free medium compared to the 10% serum-supplemented medium(p < 0.05). The functional differentiation of the primary culture was examined by observing multicellular domes of the confluent monolayer, which is indicative of transepithelial solute transport. The dome formation by the proximal tubule cultures occurred at a higher frequency in the hormonally defined, serum-free medium than in the 10% serum-supplemented medium(p < 0.05). Upon electron microscopic examination, an increased density of the brush border was observed in the hormonally defined, serum-free medium compared to the cells grown in 10% serum-supplemented medium. The activities of $Na^+$glucose cotransporter($^{14}C$-a-MG uptake), $Na^+$phosphate cotransportere($^{32}P$ uptake) and $Na^+$ transporter($^{22}Na^+$ uptake) in the brush border membrane, and of $Na^+/K^+$-ATPase($^{86}Rb$ uptake) in the basolateral membrane were significantly stimulated in the hormonally defined, serum-free medium than in 10% serum-supplemented medium(p < 0.05). In conclusion, the primary cultured proximal tubule cells grown in the hormonally defined, serum-free medium demonstrated a slower growth rate, but the functions of cell were enhanced.

• Korean Journal of Weed Science
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• v.10 no.4
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• pp.285-293
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• 1990

The response of callus growth of rice (Oryza sativa L.) cultivars which showed different responses to herbicides as seedlings was investigated to select resistant or tolerant calli. Callus growth of IR28 which was susceptible to thiobencarb (S-[(4-chlorophenyl)methyl]diethylcarbamothioate) during callus induction was not inhibited by $10^{-5}$ M and $10^{-6}$ M thiobencarb, indicating that there was a difference in tolerance among callus induction and growth, and the intact plant level. A similar result was obtained with IR31917-45-3-2-2 to butachlor [N - (buthoxymethyl) -2-chloro-N- (2, 6-diethylphenyl) acetamide]. The fresh weight of IR28 callus transferred into $10^{-5}$ M thiobencarb after treatment at $10^{-6}$ M for 30 days was not affected by the herbicide, indicating that transferring callus into gradually higher herbicide concentrations can be a useful method for selection of herbicide-tolerant cell lines.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.647-659
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• 1997

To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.4
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• pp.316-320
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• 2015

In order to improve the transformation efficiency of zoysiagrass by increasing the frequency of callus formation from mature seeds and plant regeneration, the effect of pre-treatment with sea sand was examined. Mature zoysiagrass seeds were given 10 min of swelling time before sea sand treatment using a sea sand and seed ratio of 1 : 1 and a vortex shaking speed of 6 (1,000 rpm) for 10 min. The seeds showed increased callus formation that was more than 2 times the rate in the control. In addition, plant regeneration efficiency was also increased when embryogenic callus induced from sea sand-treated mature seeds was cultured in regeneration medium. These results will be very helpful for improving the genetic transformation frequency of zoysiagrass, a recalcitrant monocot grass.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.1
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• pp.1-8
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• 2011

Molecular techniques such as genetic transformation are powerful tools that can be used for the genetic modification of warm-season grasses. The P. meyerianum with high regeneration ability was used for establishing an Agrobacterium-mediated transformation system. We investigated various factors affecting Agrobacterium infection by examining GUS gene expression of pCAMBIA1304 vector. Among various concentration of acetosyringone and betaine tested for inoculation and co-cultivation, 10 mg/L acetosyringone and 60 mg/L betaine resulted in the highest transformation frequency in terms of GUS expression. The calli of 4 species of Panicum spp. with excellent tissue culture response were inoculated with Agrobacterium under the optimal infection conditions. The high activity of GUS gene was observed in all species and hygromycin-resistant calli expressing GFP were obtained in P. meyerianum, P. longijubatum, P. stapfianum and guineagrass Noh-PL1. Co-cultivated calli were transferred onto the selection medium containing hygromycin, and the hygromycin resistant calli were selected after 3 months. Hygromycin-resistant plantlets were then successfully regenerated from the calli and grown in a greenhouse. We confirmed stable insertion of hpt gene among the hygromycin-resistant plantlets of P. meyerianum by PCR analysis.

• Journal of Life Science
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• v.19 no.6
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• pp.728-734
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• 2009

In this work, a simple, inexpensive and reproducible technique of flotation density gradient centrifugation was developed to isolate monocytes with high purity and yield from peripheral blood mononuclear cells (PBMC) using Histopaque solution, density and osmolarity of which were modified to 1.072 g/ml and 335 mOsm with phosphate-buffered saline (PBS) and sodium chloride (NaCl) solution, respectively. The average purity of monocytes was 74.75${\pm}$3.84%, with the individual purity ranging from 71.44% to 82.38%. The average yield of monocytes was 32.62${\pm}$11.16%, with the individual yields ranging from 21.02 to 53.63%. The monocytes isolated by floatation density gradient centrifugation could be successfully cultured into morphologically, phenotypically and functionally dendritic cells in vitro. In conclusion, the entire procedure seemed to be faster and more convenient, simple and cost-effective than other monocyte isolation methods, including plastic adherence and density gradient methods, and has the potential to be developed as a closed system for clinical scale generation of dendritic cells.

• Journal of Life Science
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• v.11 no.6
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• pp.603-611
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• 2001

Phytolalexins are produced in plants affected by various environmental factors such as fungal infection treatment with many chemicals and irradiation by ultraviolet light. When pepper and tobacco bel suspension cultures were grown on a basal MS medium supplemented with 2,4-D(1mg/$\ell$, benzyl adenine(0.001 mg/$\ell$) and 100$\mu$ M jasmonic acid, the production of capsidiol was observed. The total of compound found in pepper plant were around seventy and thirty of them were located intissue-specific manner. 1-propanethiol, $\alpha$-D-xylofuranoside, phenol, hexadecanonic acid ethyl tridecanoate, phytol, linoleic acid and capsidiol are those which have change the production level by treatments, such as the inoculation of Phytophthora capsici Leonian, the metalaxyl treatment and the UV-B irradiation, respectively. The content of capsidiol on inoculation of P. capsici with metalxyl suspension in soil were higher than those of P.capsici without metalaxyl. When the soil dernch of metalaxyl treatment (1$\mu\textrm{g}$/${mu}ell$)was delayed after inoculation, the content of capsidiol were higher than that of before. Irrradiated UB-B the production on capsidiol was identified only at leaf, and contents were the highest for 24 hrs incubation after 20 minutes irradiation.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.124-129
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• 2016

Endotoxin, also known as lipopolysaccharide (LPS) produced by the cell wall of gram negative bacteria can be present in any liquid or on any biomaterial. Endotoxin in blood can cause fever and inflammation. In this study, we compared bacterial endotoxin using Escherichia coli O157:H7, Klebsiella oxytoca, Salmonella Typhi, Shigella sonnei and Morganella morganii. Bacteria were cultured for use in the experiment, and diluted to $1.5{\times}10^8CFU/mL$. A check marked sensitivity confirmatory test of the Limulus amebocyte lysate (LAL) reagent was performed to examine the validity. The end point reaction to each bacteria sample was confirmed with 10 fold dilution and then the final reaction end point was confirmed by 2 fold dilution between the dilution step and the upper dilution step. According to the results, in detection of endotoxins in more than 0.015 EU/mL, E. coli O157 was 75~37.5 CFU/mL, K. oxytoca 37.5~18.75 CFU/mL, M. morganii and S. Typhi 3.75~1.875 CFU/mL, and S. sonnei 7.5~3.75 CFU/mL. The resulting value was finally ensured by a confirmation test for the inhibitory factor. Based on this study, conduct of further research on bacterial endotoxin is encouraged.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.89-94
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• 2013

The present study was undertaken to elucidate the mechanisms underlying the cholesterol-lowering effect of S-allylmercaptocysteine (SAMC) derived from aged garlic. Rat hepotocytes and HepG2 cells were used to determine the short-term effects of SAMC on [$^{14}C$] acetate incorporation into cholesterol, and several enzymatic steps. The cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and treated with 20, 40, 60 and 80 ${\mu}g/ml$ of SAMC. At concentration of 20~40 ${\mu}g/ml$, no significant cells viability effect was noted during those incubation periods. However, at a concentration 60 ${\mu}g/ml$, cell viability decreased approximately 50% compared with the control. The treatment of cells with 5, 10, 15, and 20 ${\mu}g/ml$ of SAMC resulted in a marked of [$^{14}C$]-acetate incorporation into cholesterol. At concentration of 15 ${\mu}g/ml$, the cholesterol synthesis was inhibited 79% in cells. The activities of lipogenic enzymes, fatty acid synthase (FAS), and glucose-6-phosphate dehydrogenase (G3PDH) were measured in culture hepatocytes treated with the inhibitors. The activity of FAS in cells treated with 0.95 nmol SAMC was 19% lower than that of nontreated cells, and no affected G6PDH activity, 3-hydroxy-3-methylglutaryl Co A activity was decreased at concentration dependant manner. The present study demonstrates that SAMC is effective in inhibiting cholesterol biosynthesis.

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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• pp.261-270
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• 1986

A total of 339 urethral, vaginal swab and eye discharge materials from the out-patients in the hospitals of Seoul area was microbiologically collected for the detection of Chlamydia trachomatis infection during May through August, 1985. McCoy cell culture system (MCC) and enzyme-likned immunosorbent assay (EIA) methods were employed in this study as the tools for the detection of C. trachomatis, and the detectabilities of two methods were compared. The results obtained in this study are summarized as follows: 1. The positive rate of C. trachomatis in 339 swab specimens was 18.6%, and the rate in females (20.1%) was much higher than that in males (7.1%). 2. The positive rate of C. trachomatis infection the prostitutes was the highest (24.2%), and the rate in the eye discharge specimens obtained from the new barns was 12.8%. 3. The positive rates of C. trachmoatis infection detected in the specimens from the patients with vaginitis and leucorrhea, with infertility, with cystitis and with nongonococcal urethritis were 17.2%, 21.9%. 18.0% and 7.1%, respectively. 4. The positive rate of C. trachomatis infection in 20-25 age group was 30.5%. This rate was the highest among the other age groups. 5. The positive rate of C. trachomatis infection in the randomly screened 89 swab specimens by EIA (30.3%) was much higher than the rate detected by MCC (18.6%). 6. The positive rate of C. trachomatis infection in females detected by EIA was also much higher than in males, and the 20-25 age group showed the highest positive rate as compared to the other age groups. 7. Sensitivity and specificity of EIA for the detection on C. trachomatis were 100% and 88.6%, respectively, in case that MCC was regarded as perfect method. In summarizing the above results, it is known that considerable cases with genital diseases and with eye discharges were associated with C. trachomatis, and that EIA method is recommendable for the detection of C. trachomatis especially in the specimens swabed from the genital tracts.