• Development and Reproduction
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• v.16 no.4
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• pp.353-361
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• 2012

Human embryonic stem (ES) cells are a potential source of cells for developmental studies and for a variety of applications in transplantation therapies and drug discovery. However, human ES cells are difficult to culture and maintain at a large scale, which is one of the most serious obstacles in human ES cell research. Culture of human ES cells on MEF cells after disassociation with accutase has previously been demonstrated by other research groups. Here, we confirmed that human ES cells (H9) can maintain stem cell properties when the cells are passaged as single cells under a feeder-free culture condition. Accutase-dissociated human ES cells showed normal karyotype, stem cell marker expression, and morphology. We prepared frozen stocks during the culture period, thawed two of the human ES cell stocks, and analyzed the cells after culture with the same method. Although the cells revealed normal expression of stem cell marker genes, they had abnormal karyotypes. Therefore, we suggest that accutase-dissociated single cells can be usefully expanded in a feeder-free condition but chromosomal modification should be considered in the culture after freeze-thawing.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.65-75
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• 2003

Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.19 no.1
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• pp.123-134
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• 2006

Objective : This experimental study was performed to investigate the effect of Herbe-medicine used for eye disease(Sean-tang, Jinpi-san, Tangpo-san, Copitdis Rhizoma) on Staphylococcus epidermidis keratitis. Methods : After administering various herbal eye drops on Staphylococcus epidermidis, I measured MIC and the size of inhibition zone. MIC was measured by dropping from 20 to $50{\mu}{\ell}$ according to density. Anti-bacterial potency was measured by the size of inhibition zone with change of volume under 2 days culture condition. Also continuous anti-bacterial potency of herbal eye drops was measured by the size of inhibition zone according to 2 days and 6 days culture each under the $50{\mu}{\ell}$ condition. Results : 1. MIC on Staphylococcus epidermidis in Sean-tang, Jinpi-san was 1%, $50{\mu}{\ell}$ 2. MIC on Staphylococcus epidermidis in Tangpo-san, Coptidis rhizoma was 10%, $50{\mu}{\ell}$ 3. MIC on Staphylococcus epidermidis in Cravit was 0.1%, $20{\mu}{\ell}$. 4. Under the 2 days culture condition, the size of inhibition zone on Staphylococcus epidermidis by volume for Sean-tang was 10.0mm in $50{\mu}{\ell}$, Jinpi-san was 16.0mm in $50{\mu}{\ell}$, Tangpo-san was 17.5mm in $50{\mu}{\ell}$, Coptidis rhizoma was 31.0mm in $50{\mu}{\ell}$ and Cravit was 34mm in $50{\mu}{\ell}$, showed the highest anti-bacterial potency. 5. Under the $50{\mu}{\ell}$ condition, the size of inhibition zone on Staphylococcus epidermidis by 2 and 6 culture days for Sean-tang was 47.5mm in 6days, Jinpi-san was 36.0mm in 6days, Tangpo-san was 45.0mm in 6 days and Cravit was 48.0mm in 6 days, which showed the highest anti-bacterial potency. 6. Under the $50{\mu}{\ell}$ culture condition, the size of inhibition zone on Staphylococcus epidermidis by 2 and 6 culture days for Coptidis rhizoma was 31.0mm in 2 days and 6 days, which showed the same anti-bacteria1 potency. Conclusions : The present author think that Herbe-medicine used for eye disease can be used to cure Staphylococcus epidermidis keratitis and if further study is performed, the use of the Herbe-medicine used for eye disease will be valuable and beneficial in the clinical medicines.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.3
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• pp.308-313
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• 1995

In relation to shortening generation by using embryo culture of barley immature seed, it is important to find out profitable embryo age for embryo culture and to understand the relationship among embryo and other characters. Two varieties(Olbori and Dusan #12) were cultivated under two growth conditions(15/10 and 25/15$^{\circ}C$). The embryos were aseptically excised when immature seeds were collected 9, 13, 17, 21, 25 and 29 days after heading and cultured on $B_5$ medium. On the 21st day after heading, the length of embryos from top or middle part of spike was longer than that from bottom part. Embryos from bottom part under low temperature condition had the shortest length. Shoot length, root length and root number after embryo culture were little difference among three parts of spike under high temperature condition. Under low temperature, seedlings from bottom part of spike were inferior to those from top or middle part. Length of 29-day-old embryos under low temperature condition was similar to that of 17-day-old ones under high. Under high temperature condition, the length of 17-day-old embryo had positive correlation with kernel width, shoot length, root length and root number, but that of 21-day-old one didn't have. Seventeen-day-old embryos obtained from 25/15$^{\circ}C$ growth condition seem to be efficient to shortening generation by using embryo culture.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.291-294
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• 2012

Embryonic stem cell classically cultured on feeder layer with FBS contained ES medium. Feeder-free mouse ES cell culture systems are essential to avoid the possible contamination of nonES cells. First we determined the difference between ES cell and MEF by Oct4 population. We demonstrate to culture and to induce differentiation on feeder free condition using a commercially available mouse ES cell lines.

• Mycobiology
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• v.34 no.2
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• pp.83-91
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• 2006

Characteristic growth patterns of Cordyceps militaris isolates on various media, under varying light conditions and at varying incubation periods were examined. Light was found to be the most critical single factor in determining the density, texture, and pigmentation of the mycelial culture of the fungus. However, under the light condition, the degree of pigmentation and mycelial density were found to be affected by the incubation period and type of medium. Irrespective of the variations in medium type or incubation period, there was no pigmentation of the mycelium under dark condition. Radial growth of the mycelium was faster under dark incubation rather than under light incubation. Abundant mycelial density and darkest pigmentation of C. militaris isolates were produced in nutritionally rich media like SDAY, SMAY and CZYA, suggesting that these media may fulfill all the requirements for vegetative growth of the fungus. Growth characteristics of C. militaris isolates could be easily observed by the simple agar culture method, which would be useful to characterize the phenotypic characteristics of large number of pure cultures of the fungus under given conditions of growth factors such as medium, light and temperature.

• Korean Journal of Weed Science
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• v.17 no.1
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• pp.24-30
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• 1997

The influence of nutrients on the phytotoxicity of herbicides (bensulfuron-methyl, pyrazosulfuron-ethyl, imazosulfuron, dimepiperate, and molinate) was investigated in controlled-environment growth chamber with direct-seeded rice (Oryza sativa L. cv. Dongjin). The phytotoxicity of bensulfuron-methyl, pyrazosulfuron-ethyl, and imazosulfuron for rice was greater in nutrient culture than in no-nutrient condition. The root growth of rice applied with these herbicides was more inhibited than the shoot growth. The most severe inhibition was obtained with pyrazosulfuron-ethyl application. The growth inhibition of rice applied by dimepiperate was increased under no-nutrient culture condition. Dimepiperate suppressed more remarkably shoot growth than root. Especially the shoot elongation was much more inhibited than the others. The shoot growth inhibition in rice applied by molinate was severer than the root. The shoot growth was reduced under nutrient culture condition, while the root growth was reduced under no-nutrient culture.

• KSBB Journal
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• v.16 no.6
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• pp.568-572
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• 2001

Light is one of the most important environmental factors controlling plant physiology. The human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was produced from cell suspension cultures of transgenic tobacco under different light conditions (24 hr light, 18 hr light/dark cycle, dark). Under 24 hr light condition, cell growth was best and dry cell weight reached 14.4 g/L. Light did not influenced the secretion of total proteins. However, in the dark condition, the ratio of secreted total protein/dry cell weight was 1.5 fold higher than those of ethel conditions. Production of hGM-CSF was highest with 18 hr light condition and reached 496.5 ug/L. In addition, the content of hGM-CSf in secreted total proteins was 1.8 fold higher than that of 24 hr light condition, which is beneficial for the purificationof the protein.

• Journal of the Korean housing association
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• v.27 no.5
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• pp.55-63
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• 2016

This study intended to establish a direction for the revitalization of youth culture centers by examining the current status of youth centers operated in Daejeon and how they have been used. Four youth centers in Daejeon were visited and a survey was conducted targeting 180 people using the centers. Frequency, Mean, and ${\chi}^2$ analysis were performed by using SPSS statistics package, and major research outcomes are as follows; There were 6 youth culture centers in Daejeon and showed a lower construction rate of 7.4% on the basis of 81 up, myun and dong. Most youth centers were small, around $300-500m^2$ in total floor area, and accommodated 100-200 people, and the space comprised multipurpose hall, cafeteria, open space, club space, multimedia space, information service room, and guide booth. There were no sports spaces among the target facilities. So, it has created the need for physical activity space to promote health & development. Most users visited centers with their friends and simply to use the facilities, and the satisfaction with the facilities was relatively high at 4.32. The role of the local community and financial support of the government is required to activate the youth culture. Also, for the role of youth culture center, it was suggested that the youth culture center should develop program which corresponds with the level of the youth and boast the interest of the youth.

• Journal of Plant Biology
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• v.39 no.1
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• pp.63-69
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• 1996

A hairy root clone of Panax ginseng C.A. Meyer, HRB-15 was cultured iu various conditions with 3 L bubble type bioreactor to enhance both growth and ginsenoside production. The hairy roots were more rapidly grown under the dark condition than under the light condition. However, total amount of ginsenoside of hairy roots cultured under the light for 30 days increased 2 folds as compared with the dark condition and was 1.10% based on 6 ginsenosides. Especially, ginsenoside-Re was significantly increased and some ginsenosides except for ginsenoside-Re was slightly reduced. Also, the growth of hairy roots decreased about 30% as compared with the dark condition. In contrast, addition of sodium acetate led to decreased production of ginsenoside and growth of hairy roots under light condition. The influence of potassium dihydrogenphosphate concentration was examined in MS medium and a 1.25 mM concentration was found to be the most appropriate for growth and ginsenoside production under light condition. Two-step process of hairy roots culture with yeast elicitation or without ammonia in culture medium was developed to enhance growth and giusenoside synthesis. $50\;\mu\textrm{g}$ of yeast elicitor per g of fresh weight showed a synergistic effect on the ginsenoside synthesis of hairy roots on 20 days after culture. At that time, the content of total ginsenoside was 1.15%, while the growth of hairy roots decreased 21 % as compared with the dark condition. In addition, when elimination of ammonia on 20 days after culture, the content of total ginsenoside was 1.26% with significant increment of ginsenoside-Rd (0.27%) in addition to ginsenoside-Re and the growth of hairy roots decreased 10% as compared with the dark condition. In this system, we have demonstrated a unique two-step process of hairy root cultures to maximize biomass and secondary metabolites. It has found possibility to enhance ginsenosides production by growing hairy roots in this method.