• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.125-133
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• 2001

The goal of this paper was to examine the qualify zygote-acquiring method for in-vitro culture and the in-vitro culture method of the acquired zygote from a technological perspective. We have reported the results on the introduction of foreign DNAs using the described culturing method. After performing in-vitro and surrogate eggshell culture on a zygote acquired from the abdomen of a hen, 25.8% hatchability was acquired. After microinjecting foreign DNAs into the acquired zygote and performing in-vitro and surrogate eggshell culture using the same method, 13.1∼11.7% hatchability was acquired. Having compared the developments of the control subjects and the experimental subjects, the viability of the experimental subjects on the 4∼5th day of culturing was much lower compared to that of the control subjects. This is a result that shows that the microinjection process of foreign DNAs might have a negative effect on the existence of the embryo; therefore, various technical attempts should be made to minimize such negative effects. Having microinjected foreign DNAs into the zygote of a hen to produce transgenic chickens, 3 transgenic founders were Produced and 70 G1 progeny were produced as a result of the progeny test that had been performed to the present.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.80-86
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• 2017

One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.

• International Journal of Oral Biology
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• v.40 no.3
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• pp.143-146
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• 2015

The purpose of this study was to compare the effect of different methods of hand washing by counting the number of bacteria on the hand surface. Eighteen clinicians were chosen and divided into three groups, consisting of six clinicians each. Culturing of the right raw palms of all individuals was performed. Individuals in the control group washed hands for 5 seconds with antimicrobial soap. Group 1 washed their hands for 10 seconds with antimicrobial soap. Group 2 washed with an instant alcohol-based hand sanitizer. After the respective washes, re-culturing of the right raw palm was done for each member of all groups. The colony-forming units (CFU) were calculated at each time point, and the reduction rate of CFU among the three groups were statistically evaluated using student t-test. All groups showed a significant decrease in CFU, according to the time applied (P<0.01). In addition, the reduction rate of CFU between the groups were statistically evaluated with ANOVA (P<0.01). It showed statistically difference between the control group and group 1, control group and group 2. The present study confirmed that the hand washing method with antimicrobial soap for 10 seconds and hand sanitizer, including alcohol, were excellent for decreasing the number of bacteria on the hand surface.

• IMMUNE NETWORK
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• v.14 no.1
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• pp.54-65
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• 2014

Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent, with the ability to differentiate into different cell types. Additionally, the immunomodulatory activity of MSCs can downregulate inflammatory responses. The use of MSCs to repair injured tissues and treat inflammation, including in neuroimmune diseases, has been extensively explored. Although MSCs have emerged as a promising resource for the treatment of neuroimmune diseases, attempts to define the molecular properties of MSCs have been limited by the heterogeneity of MSC populations. We recently developed a new method, the subfractionation culturing method, to isolate homogeneous human clonal MSCs (hcMSCs). The hcMSCs were able to differentiate into fat, cartilage, bone, neuroglia, and liver cell types. In this study, to better understand the properties of neurally differentiated MSCs, gene expression in highly homogeneous hcMSCs was analyzed. Neural differentiation of hcMSCs was induced for 14 days. Thereafter, RNA and genomic DNA was isolated and subjected to microarray analysis and DNA methylation array analysis, respectively. We correlated the transcriptome of hcMSCs during neural differentiation with the DNA methylation status. Here, we describe and discuss the gene expression profile of neurally differentiated hcMSCs. These findings will expand our understanding of the molecular properties of MSCs and contribute to the development of cell therapy for neuroimmune diseases.

• Journal of Environmental Science International
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• v.23 no.5
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• pp.771-780
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• 2014

Tetragonia tetragonides is a medicinal plant native to ocean sand soil of southern provinces and has significant effects on the prevention and curing of gastroenteric disorders. Despite of its popularity, supply of the plant has never met the level of demand because of the absence of an adequate culturing method. The present study, thereby, was conducted for classifying the plants with geographically different characteristics, studying growth habits, developing a new culturing method and establishing a large scale propagation system of selected superior individual plants. The study was also aimed for revealing optimum conditions for seed treatment, fertilization, and efficient culturing system and thereby, for utilizing the plant as a new income source for rural communities. The seed was elongated with size of 2.6 mm (width) ${\times}$ 1.8 mm (length). No difference in seed size was observed depending on different inhabitate. Each flower produced about 4.5~4.8 seeds. Germination rate was high for seeds matured for 40 days after fertilization, but deceased to 50% for seeds matured only for 20 or 30 days. Seed dormancy lasted 6 months and seed storage at humid $5^{\circ}C$ facilitated germination. Mechanical obstruct of seed germination was due to seed coat and removal of seed coat enhanced the germination rate. Optimum temp. for seed storage was $5^{\circ}C$, and high germination rate was maintained for 350 days. However, for stratification condition or at room temperature, germination was significantly reduced as storage time increased Optimum treatment of plant growth regulators was soaking in $GA_3$ 250 mg/L for 1 hr. The priming treatment with 50 mM $Ca(NO_3)_2$ at $20^{\circ}C$ for two days improved the seed germination with 10% compared to non-treated control. The treatment of 20% NaOCl for 3 hr. improved the seed germination rate up to 10% and 1 day ahead.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.1
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• pp.44-49
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• 2003

In vitro culture of panicle has been the method to accumulate starch and protein in immature grains by providing nutrients after florets crossed between remote genotypes artificially. Grain filling means embryo development and sucrose translocation from photosynthetic source, and starch manufacture in endosperm. The concentrations of sucrose used to culture immature rice panicle were 5, 10, 15, 20% and glutamine was 20 mM. When immature rice panicles at 5 days after flowering were cultured in distilled water with different concentrations of sucrose, glutamine 20 mM and MS medium with different concentrations of sucrose, glutamine 20 mM for 30 days the later was effective on grain filling. The optimal concentration of sucrose on grain filling in vitro culture of rice panicle was 10% and the weight of grain cultured was 10.14 mg that was corresponded to 57% of intact plant. In the method of treating plant growth regulators, the culture of immature rice panicle adding in MS medium with Kinetin, IAA, $\textrm{GA}_3$ were effective on grain filling than the culturing of immature rice panicle after submerging in solutions of Kinetin, IAA, $\textrm{GA}_3$ for 1day. When immature rice panicle was cultured in MS medium with sucrose 10% and Kinetin 46.47 $\mu$M it was effective on grain filling, respectively. The weight of grain cultured was 13.1mg that was corresponded to 75% of intact and germination rate was 51 %. When immature rice panicles were cultured in medium with different concentrations combined with Kinetin 4.65, 46.47, 464.7 $\mu\textrm{M}$, IAA 5.71, 57.08, 570.80 $\mu\textrm{M}$ for 30 days and in medium with IAA 5.71, 57.08, 570.80 $\mu\textrm{M}$ for 15 days after culturing in medium with Kinetin 4.65, 46.47, 464.70 $\mu\textrm{M}$ for 15 days the effect on grain filling was similar.

• Journal of Environmental Health Sciences
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• v.45 no.5
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• pp.511-519
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• 2019

Objectives: The standard method for the enumeration of environmental Legionella is culturing, which has several disadvantages, including long incubation and poor sensitivity. The purpose of this study is to demonstrate the usefulness of real-time PCR and to improve the standard method. Methods: In 200 environmental water samples, a real-time PCR and culture were conducted to detect and quantify Legionella. Using with the results of the survey, we compared the real-time PCR with the culture. Results: Each real-time PCR assay had 100% specificity and excellent sensitivity (5 GU/reaction). In the culture, 36 samples were positive and 164 samples were negative. Based on the results of the culture, real-time PCR showed a high negative predictive value of 99%, 35 samples were true positive, 105 samples were true negative, 59 samples were false positive and one sample was a false negative. Quantitative analysis of the two methods indicated a weak linear correlation ($r^2=0.29$, $r^2=0.61$, respectively). Conclusions: Although it is difficult to directly apply quantitative analysis results of real-time PCR in the enumeration of environmental Legionella, it can be used as a complementary means of culturing to rapidly screen negative samples and to improve the accuracy of diagnosis.

• Journal of Mushroom
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• v.2 no.1
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• pp.15-20
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• 2004

Pleurotus ostreatus, the oyster mushroom, is one of the most widely cultivated and important edible mushrooms in the world. In order to study the developmental process of P. ostreatus and its regulatory mechanism, a new culturing method needs to be established for inducing the fruiting body and sporulation in the laboratory. In this study, we have examined whether the fruiting body of P. ostreatus can be formed on the plastic petri dish which are commonly used for cell culture in the laboratory. The strain was cultured on $60{\times}15mm$ plastic petri dish with potato dextrose agar media at $28^{\circ}C$ for mycelial growth and then at $18^{\circ}C$ for the formation of primordia and fruiting bodies within plant growth chamber. The development of primordia into fruiting bodies was achieved on cultured dishes under air ventilation. At the primordia stage, the normal formation of fruiting body was blocked by sealing the plastic dish with parafilm. The periods requiring for the formation of primordia and fruiting bodies were examined on the dish culture. About 96% and 76% of cultured samples formed primordia and fruiting bodies under the optimal conditions during ten weeks of culture, respectively. These culturing periods, however, were changed by the mechanical injury treatment to mycelia. As other factors affecting the fruiting body formation, the effects of light and cold shock have been tested. No fruiting formation was observed on the cultured dishes under the dark. The cold shock treatment by storing cultured dishes for one day at $4^{\circ}C$ did not have any significant effects in the fruiting body formation. Spores of fruiting bodies acquired from the petri dishes could be germinated on culture media at $28^{\circ}C$. These results suggest that the fruiting bodies of P. ostreatus can be formed on the experimental petri dish and this dish-culturing method is useful for understanding of the developmental process of P. ostreatus in the laboratory. Furthermore, the dish-culturing method is able to shorten the life cycle of P. ostreatus without requiring large area and expensive device.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.31-35
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• 2004

In order to develop an efficient micropropagation technique for an endangered species, Stellera rosea N., stem node cultures were conducted on MS medium supplemented with cytokinins. Generally, BA was better than zeatin on shoot proliferation from stem nodes, whereas zeatin showed more effective on shoot elongation. In vitro rooting of shoots was achieved by application of an auxin pre-culturing method. Overall rooting rate was relatively low and differed depending on the culture period. Pre-culturing of shoots for 15 days at 1.0mg/L IBA revealed a slightly better rooting efficiency reaching 30％ rooting rate than NAA. Root induction rate by NAA also varied with concentration of NAA and culture periods. Total 51％ of the rooted plantlets survived on artificial soil mixture and grew normally without any distinct morphological variation. The results suggest that the endangered Stetllera plants are propagated via in vitro culture system, but still need to more study for the improvement of rooting and acclimatization of the plantlets in soil.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.56-60
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• 1976

In order to find effective seed collection method from cultivated Porphyra, benthic diatom elimination, culturing conditions of Conchocelis, liberation of conchospores and treatment of the fronds to obtain monospores have been studied. Elimination of benthic diatoms from Porphyra fronds is successfully performed by careful brushing the fronds in sea water and freshwater alternatively. For the culturing of Conchocelis Schleiber's solution enriched with only soil extracts, vitamins and Fe-EDTA was satisfactory. Growth under 16 hours illumination is 1.29 times faster than those under 10 hours illumination. When the culturing water was airated the growth was $1.41\~l.50$ times faster than the growth in stagnant water. Total amount of conchospores liberated from Conchocelis which has been cultured under airation was much more than those of conchospores under stagnant condition. Effective liberation of monospores was observed in the fronds which have been dried in air for 6 hours $(21.23\~24.19\%\;water\;content)$.