• FLOWER RESEARCH JOURNAL
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• v.18 no.2
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• pp.83-86
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• 2010

This study was conducted to develop in vitro propagation techniques of new cultivar, 'Greenstar', bred by Highland Agriculture Research Center. The multiple shoot induction and plant growth of in vitro plant were analyzed by MS media concentration (1/2 MS, 1 MS and 2 MS medium), plant growth regulators and its proper concentration; CPPU [Forchlorfenuron(N-(2-chloro-4-pridyl)-3-phenylurea) (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), thidiazuron [(TDZ), (0.01, 0.1, 0.5 and $1.0mg{\cdot}L^{-1}$), zeatin (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$), and BA [6-benzylaminiopurine(BA), (0, 0.5, 1.0 and $2.0mg{\cdot}L^{-1}$)] in MS (3% sugar and 0.8% agar with pH 5.7) media. The highest number of induced shoots, leaves and roots were shown in 1/2 MS medium concentration. On the 1/2 MS medium, shoot numbers, shoot length, leaf numbers and root numbers were 11.0, 1.9 cm, 24.7, and 8.0, respectively. On the absence of CPPU in the 1/2 MS medium, shoot length and root numbers was greater than CPPU treatment, but the highest number of shoots was induced by the $2.0mg{\cdot}L^{-1}$ of CPPU concentration in 1/2MS medium. TDZ, zeatin, and BA treatments were not effective on the induction of multiple shoot in vitro culture. As a result, in vitro culture of new Saxifraga fortunei, 'Greenstar' with $2.0mg{\cdot}L^{-1}$ of CPPU in 1/2 MS medium was most effective for the rapid multiplication.

• Journal of Plant Biotechnology
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• v.46 no.2
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• pp.119-126
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• 2019

This study was conducted to investigate the optimal culture method for gametophyte and sporophyte propagation in Coniogramme japonica (Thunb.) Diels, which can be used in various fields. The propagation of prothallus were cultured in 1/4 - 1 Murashige and Skoog medium and Knop medium for 10 weeks. The results indicated that the fresh weight of prothallus was the highest (14.5 g) in 1MS medium. Subsequently, various concentrations of sucrose, activated charcoal and nitrogen source were also added to 1MS medium and cultured for 8 weeks. The results provided that the sucrose concentration was 3% and the fresh weight of prothallus was the highest 10.8 g. According to the concentration in the range of 8.8 ~ 10.8 g, in the case of activated charcoal, the four treatments showed no significant difference. The nitrogen source was added at a concentration of 30, 60 and 120 mM with the ratio of ${NH_4}^+:{NO_3}^-$ being 1 : 2. As a result, the fresh weight of all treatments increased to similar level but there was no significant difference. We investigated sporophyte formation according to soil type and the highest number of sporophytes at 228.0 was formed in soils mixed with horticultural substrate and decomposed granite at 2 : 1 (v : v). On the other hand, sporophyte was not formed in soils containing peatmoss except for the one with peatmoss and decomposed granite at 2 : 1 (v : v).

• Development and Reproduction
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• v.12 no.3
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• pp.221-229
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• 2008

For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.

• Tuberculosis and Respiratory Diseases
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• v.66 no.3
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• pp.178-185
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• 2009

Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

• Horticultural Science & Technology
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• v.18 no.6
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• pp.834-838
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• 2000

As an endeavor to establish a micropropagation system for Epimedium koreanum Nakai., this study was carried out to define methods to disinfect its explants and media for callus induction, proliferation and plant regeneration. The lowest infection rates by fungi or bacteria on apical and axillary bud explants of rhizome were observed when they were immerged in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 30 min, but leaf explants were seldom infected with fungi or bacteria by this disinfectant method. The highest rate of plantlet formation was obtained from the explants disinfected in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 60 min for tip buds and in 0.1 % $AgNO_3$ solution for 30 min for axillary buds of rhizome. Induction rate of callus was the highest from the explants disinfectd in 0.3% NaOCl solution for 20 min after soaked in 0.2% $AgNO_3$ solution for 15 min. Callus growth was proper in a modified 1/2 MS medium including half strength of $NH_4NO_3$ with $0.02-0.2mg{\cdot}L^{-1}$ BA and $2.0mg{\cdot}L^{-1}$ NAA. Low rate of plantlet regeneration was obtained in 1/2 UM or 1/2 White medium with $2.0mg{\cdot}L^{-1}$ BA and $0.2mg{\cdot}L^{-1}$ AA.

• Korean Journal of Veterinary Pathology
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• v.6 no.2
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• pp.101-104
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• 2002

닭의 전신장기에 림프종 발생이 특정인 마렉병(Marek's disease; MD)은 림프구성 백혈병(Lymphoid leukosis; LL)과 병리학적 소견이 매우 유사하여 감별이 요구된다. 마렉병 바이러스(Marek's disease virus; MDV)는 질병초기에서 후기에 이르기까지 모낭상피세포에 세포용해성 감염을 지속적으로 일으킨다. 세포용해성 감염이 있는 모낭상피세포는 변성내지 괴사되어 있고 핵내봉입체가 관찰된다. 또한 세포용해성 감염이 있는 모낭상피세포 바로 밑의 진피와 깃털 수질(feather pulp)내의 혈관주위에 림프구 침윤이 관찰된다. 이러한 피부병변은 MD의 특징적인 병변들로서 LL과 감별할 수 있는 결정적인 단서이다. 본 고에서는 MD에 관한 전반적인 것과 특히 MD 진단을 위한 피부병변의 유용성에 대해서 자세히 논하고자 한다. 양계산업에 문제를 야기하고 있는 림프구 증식성 종양은 크게 마렉병(Marek’s disease; MD)과 백혈병(Lymphoid leukosis; LL)이 있다. 이들 질환의 원인체들이 분리되기 전까지는 이들 질병들은 발병부위에 따라 질병명을 붙였다. 즉, 내부장기냐 근육에 발생한 것은 visceral lymphomatosis, 피부에 발생한 것은 skin leukosis, 말초신경의 병변은 poly­n neuritis, neuritis, neurolymphomatosis gallinarum, range paralyis로 불리었다. 또한 눈에 발생한 것은 blindness, gray eye, iritis, uveitis, ocular lymphomatosis라고 불리었다. 1961년에 Biggs는 이러한 림프구 증식성 질병을 마렉병과 백혈병 으로 분류하자는 제안을 하였고, 드디어 1960년대 중반에 림프구 증식성 병변 중의 일부에서 herpesvirus가 분리되어서 Biggs가 제안한 병명인 마렉병을 본격적으로 사용하게 되었다. MD는 마렉병 바이러스(Marek’s disease virus; MDV)가 원 인제로서 닭에 전염성이 강한 염증성에서 종양성의 병변을 내부장기, 피부, 근육, 안구, 중추신경계, 말초신경계 등에 일 으킨다. MDV는 림프종을 유발하기 때문에 처음에는 사람에서 림프종을 유발하는 Epstein-Bar 바이러스와 관련이 있을 것으로 생각되어 gamma-herpesvirus로 분류되었지만, MDV의 게놈 구조와 세포배양에서 빠르게 성장한다는 점 때문에 지금은 alpha-herpesvirus로 재분류 되었다. MDV는 바이러스 중화시험과 한천 겔 침강법에 의해서 3개의 혈청형으로 분류 된다. 혈청형 1은 종양원성 바이러스와 종양원성 바이러스의 계대배양에 의한 약독주가 있다. 혈청형 2는 자연적으로 발 생하는 비종양원성 닭의 MDV이고, 혈정형 3은 바종양원성 칠면조 herpesvirus (HVT)이다. 림프종을 유발하는 MDV 감염은 4개의 과정, 즉 초기 세포용해성 감염, 잠복감염, 후기 세포용해성 감염, 마지막으로 종양화로 나눌 수 있다. 감염의 경로를 보면, 흡입된 MDV는 폐의 대식세포에 감염한 후 전선 장기로 MDV를 전파 시킨다. 특히, 흉선, F냥, 비장 등의 림프구에 초기 세포용해성 감염을 일으키는데, B 림프구가 주로 감염된다. 세포용해성 감염음 방어하기 위해 몰려든 T 림프구가 활성화가 되면, T 세포도 감염되게 된다. 잠복감염은 여러 가지 사이토카인 등 을 포함한 면역반응에 의해서 일어나며, 이 때 잠복감염된 세포는 특히 혈중의 T 세포라고 한다. 혈중 MDV 감염세포 는 피부 모낭상피세포, 선장 등 상피세포 유래의 조직에 MDV를 전파 시켜서 이들 조직에서 후기 세포용해성 감염이 일어나게 한다. 후기 세포용해성 감염을 유발하는 것은 이러한 MDV가 감염된 혈중 림프구라는 증거는 혈중의 세포 외에는 감염성 바이러스가 없기 때문이다. 후기 세포용해성 감염이 있는 후 육안적 혹은 현미경적으로 검출이 가능한 림프종이 여러 장기에서 관찰된다. MDV가 감염되면 병변 발생부위에 따라서 다양한 임상증상이 발생한다. 즉 내부장기에 병변이 있을 경우는 침울, 체중감소, 산란율 저하 등이 발생하며, 신경계는 발생부위별 신경증상이 일어난다. 이러한 장기나 조직에서는 육안적으로 백색에서 회백색의 종양성 병변이 관찰된다. 말초신경에 병변이 발생한 경우에는 특히 좌골신경 및 신경총에서 호발하는데, 이들 조직은 편측성 혹은 양측성으로 종창되어 있다. 안구는 간혹 육안적으로 식별이 가능한 홍채 퇴색 및 증식 병변이 관찰된다. 피부형 MD는 특히 육용계에서 문제가 되고 있으며, 산란계의 내장형 MD가 발생한 경우에도 피부를 자세히 설펴보면 피부형 MD를 간혹 관찰할 수 있다. 피부형 MD는 모낭주위에 원형의 결절형태로 발생하는데, 이들 병변이 커지면 바로 옆의 병변과 융합 되어 큰 결절을 형성하기도 한다. (중략)

• Korean Journal of Plant Taxonomy
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• v.46 no.2
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• pp.256-266
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• 2016

A total of 13,000 individuals of Dendrobium moniliforme (L.) Sw. artificially propagated in laboratories and greenhouses were restored in their natural habitat of Bogildo Island, Wandogun, in the southern part of Korea in June of 2013. The growing conditions of the individuals were monitored for two years. The parental individuals for the restoration were obtained from a wild population in southern Korea, from which seeds were produced via artificial crossings. These seeds were germinated and cultivated in growing media and two-year-old plants were then grown in greenhouse beds. The genetic diversity among the propagated individuals was confirmed by examining DNA sequences of five regions of the chloroplast genome and the nuclear ITS region. The diversity values were as high as the average values of natural populations. All propagated individuals were transplanted into two different sites on Bogildo by research teams with local residents and national park rangers. After restoration, we counted and measured the surviving individuals, vegetative propagated stems, and growth rates in June of both 2014 and 2015. There was no human interference, and 97% of the individuals survived. The number of propagules increased by 227% in two years. In contrast, the average length of the stems decreased during the period. In addition, different survival and propagation rates were recorded depending on the host plants and the restored sites. The shaded sides of rock cliffs and the bark of Quercus salicina showed the best propagation rates, followed by the bark of Camellia japonica. A few individuals of D. moniliforme successfully flowered, pollinated, and fruited after restoration. Overall, our monitoring data over two years indicate that the restored individuals were well adapted and vigorously propagated at the restored sites. In order to prevent human disturbance of the restored sites, a CCTV monitoring system powered by a solar panel was installed after the restoration. In addition, a human surveillance system is operated by national park rangers with local residents.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.2
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• pp.243-254
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• 2002

For the purpose of evaluating the biocompatability of 3 kinds of metallic materials frequently used in pediatric dentistry (stainless steel crown, orthodontic band, orthodontic wire), cellular and molecular studies, including cell growth and proliferation, screening of cell death with determination of types whether necrosis or apoptosis and changes in expressions of related signaling molecules were examined, using cultured human gingival fibroblasts (HGF-1), HGF-1 was cultured in Dulbecco's modified Eagle's medium. among which the 3rd to 6th generations of HGF-1 were used. The specimen were divided into stainless steel crown (R), band (B) and wire (W). The immunocytochemical study was done for the detection of anti-PCNA (proliferating cell nuclear antigen) labeling. With extracted protein, western blot was done for the detection of ERK1/2, JNK, and p38, using individual antibodies. Cultured cells proliferated, remarkably till 7 day and slightly at 11 day. There was no statistical significance in the counts of proliferating HGF-1 between control and experimental groups (p>0.05). Relative growth rates were no statistically significant difference between control and experimental groups (p>0.05). PCNA labeling indexes showing similar patterns in control and experimental groups. The expressions of ERK1 and ERK2, p38 were similar in control and experimental groups. The expression of JNK increased at 1st day, slightly decreased at 4th day and markedly increased at 7th and 11 day. Although the patterns of control and experimental groups were similar, the increased expressions of JNK at late period suggest a possible stress due to inhibited cell growth and proliferation, and worse culture condition. Conclusively, the 3 kinds of metal specimens used in this study did not induce cellular and molecular hazards during short term culture of HGF-1. But, for the better clinical stability, the establishment of long period culture and animal experiment was thought necessary.

• Journal of Life Science
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• v.28 no.10
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• pp.1201-1211
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• 2018

The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.179-183
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• 2002

This study was carried out to investigate the influence of medium composition for in vitro mass propagation of Lycoris squamigera Max. After the disks of short stems, segments of leaf within bulb and scale were cultured on MS basal medium supplemented with various plant growth regulators, they were examined for the extent of callus formation, shoot and root regeneration. In the culture of stem disks, adventitious shoots were regenerated from the basal tissue of bulb scales, and combined medium of 1.0 mg/L 2,4-D or NAA＋2.0 mg/L BA or kinetin showed the the best response and 4∼6 shoots per explant formed. In the culture of leaf segments within bulbs, both MS medium supplemented with 1.0 mg/L NAA＋2.0 mg/L TDZ and with 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA were produced callus profusely on the base of leaf tissue and 3∼6 shoots were regenerated per explant. In the scale segments culture, calli were produced on the basal tissue on medium with 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA. The best result were shown on MS medium with 1.0 mg/L NAA＋2.0 mg/L TDZ, and 1.0 mg/L 2,4-D＋1.0∼2.0 mg/L BA. Maximum number of regenerated shoots was up to 10∼12. Adventitious root formation from explants were formed profusely on MS medium with 1.0 mg/L NAA＋2.0 mg/L kinetin. The most desirable method for mass propagation of plantlets was the shoot regeneration from scale segments then subsequently subcultured on medium for rooting.