• Title/Summary/Keyword: in-vitro

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Transduction of eGFP Gene to Human Embryonic Stem Cells and Their Characterization (인간 배아줄기세포로의 eGFP 유전자 도입 및 특성 분석)

  • Kim, Yoon-Young;Ku, Seung-Yup;Park, Yong-Bin;Oh, Sun-Kyung;Moon, Shin-Yong;Choi, Young-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.283-292
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    • 2009
  • Objective: Human embryonic stem cells (hESCs) can proliferate indefinitely and differentiate into all kinds of cell types in vitro. Therefore, hESCs can be used as a cell source for cell-based therapy. Transduction of foreign genes to hESCs could be useful for tracing differentiation processes of hESCs and elucidation of gene function. Thus, we tried to introduce enhanced green fluorescent protein (eGFP) gene to hESCs, XX and XY cell lines in this study. Methods: Lentivirus containing eGFP was packaged in 293T cells and applied to hESCs to transduce eGFP. Expression of transduced eGFP was evaluated under the fluorescence microscope and eGFP positive population was analyzed by FACS. Expression of undifferentiation state markers such as Oct4, Nanog, SSEA4 and Tra-1-81 was examined by RT-PCR and/or immunofluorescence in eGFP-hESCs after transduction. In addition, the ability of eGFP-hESCs to form embryoid bodies (EBs) was tested. Results: eGFP was successfully transduced to hESCs by lentivirus. eGFP expression was stably maintained up to more than 40 passages. eGFP-hESCs retained expression patterns of undifferentiation state markers after transduction. Interestingly, disappearance of transduced eGFP was notably observed during spontaneous differentiation of eGFP-hESCs. Conclusion: We established eGFP expressing hESC lines using lentivirus and showed the maintenance of undifferentiation characteristics of these eGFP-hESCs. This reporter-containing hESCs could be useful for tracing the processes of differentiation of hESCs and other studies.

Effects of n-Hexane Fraction of Angelica acutiloba on Antioxidative System and Lipid peroxidation in Ethanol-Induced Hepatotoxicity of rats (일당귀 n-hexane분획이 에탄올을 투여한 흰쥐의 항산화계 및 지질과산화에 미치는 영향)

  • Choo Myung-Hi;Choi Hyun-Suk;Seo Young-Nam;Lee Myung-Yul
    • Food Science and Preservation
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    • v.11 no.3
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    • pp.364-372
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    • 2004
  • To investigate antioxidative effects of n-hexane fraction of Angelica acutiloba on the ethanol-induced hepatotoxicity of rats, Sprague-Dawley rats weighing 100 $\pm$ 20 g were divided into 5 groups; normal group(NOR), ethanol(10 mL/kg, 35$\%$) treated group(CON), n-hexane fraction of Angelica acutiloba 70 mg/kg treated group(Al), n-hexane fraction of Angelica acutiloba 70 mg/kg and ethanol treated group(A2) and n-hexane fraction of Angelica acutiloba 140 mg/kg and ethanol treated group(A3), respectively. The antioxidative activities of ethanol extract of Angelica acutiloba in vitro were decreased in order of n-hexane > ethylacetate > chlorofonn > n-butanol (>) water fraction. The growth rate and feed efficiency rate decreased by ethanol were gradually increased to the adjacent level of the normal group by administering n-hexane fraction of Angelica acutiloba. It was also observed that the activities of SOD of liver, ALT and AST of serum increased by ethanol were markedly decreased in n-hexane fraction of Angelica acutiloba administered group, and not in activites of XO, catalase, as compared with the control group. The depleted content of GSH by ethanol was increased adjacent to normal level by administering n-hexane fraction of Angelica acutiloba. as a dose-dependent manner. These results suggested that n-hexane fraction of Angelica acutiloba has a possible protective effect on the ethanol-induced hepatotoxicity of rats.

Antifungal and Antioxidant Activities of Extracts from Leaves and Flowers of Camellia japonica L. (동백나무 잎과 꽃 추출물의 항미생물 활성 및 항산화 효과)

  • Lee, Sook-Young;Hwang, Eun-Ju;Kim, Gi-Hae;Choi, Young-Bok;Lim, Chae-Young;Kim, Sun-Min
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.93-100
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    • 2005
  • This research was performed to investigate the possibilities of industrial usage of camellia (Camellia japonica L.) by examining the antioxidant and antimicrobial effects of methanol extract with different sections. Content of total phenolics, DPPH radical scavenging activities and antibacterial activity of young leaf, mature leaf, flower bud, flower, bark, and seed of camellia were compared in vitro experimental models. Total phenolics was contained the higher in young leaf (74.62 mg), flower bud (65.02 mg) and flower (62.42 mg) but less than 20.95 mg per 100 g of dry weight in other parts of Camellia japonica L. And effects of antioxidant measured by DPPH radical scavenger activity ($RC_{50}$, reduce concentration 50%), was shown higher $7.16{\sim}18.14\;{\mu}g/m{\ell}$ in methanol extract of young leaf, flower bud and flower than $61.23\;{\mu}g/m{\ell}$ of BHT as a chemical oxidant. Also, the antimicrobial activity of Camellia japonica L. extracts determined using a paper disc method against food-borne pathogen and food spoilage bacteria, the young leaves extracts showed the most active antimicrobial activity against 7 kinds of harmful microorganisms. Flower bud extracts showed the highest antibacterial activity against P. aeruginosa and Enterobacter spp. C1036. In addition, the minimum inhibitory concentration (MIC) of young leaf extract against B. subtillis,S. fradiae,S. aureus,E. coli,P. aeruginosa, Enterobacter spp. C1036, and S. typhimurium were revealed 1 to 15 ${\mu}g/m{\ell}$. As a result, antimicrobial activity of camellia extracts was shown higher gram positive bacteria than gram negative bacteria.

Biological Activity of Ixeris dentata Nakai juice Extracts (씀바귀(Ixeris dentata Nakai) 생즙 추출물의 생리활성)

  • 김명조;김주성;강원희;조미애;함승시;정동명
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.924-930
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    • 2002
  • Ixeris dentata extracts exllibited antimicrobial activity against some bacteria and fungi. Also EtOH extracts showed strong antioxidant activity and RC$_{50}$ value was 28 $\mu\textrm{g}$/mL. The inhibitory effect of Ixeris dentata on the mutagenicity in Salmonella and cytotoxicity on cancer cell were studied. Ixeris dentata extracts showed anti-mutagenic effects of 78.83 and 75.96% on B(a)P in S. typhimurium TA98 and Th100, respectively. These extracts showed 78.72% antimutagenicity on TA100 against MNNG. The Ixeris dentata extract with strong antimutagenic activities was further fractionated by hexane, ethyl acetate, butanol and water. Butanol fraction was found to be highest in antimutagenic activity against MNNG than the other fractions. Butanol fraction of Ixreis dentate revealed the highest cytotoxicity against AS49 human lung carcinoma cells in which cell growth was inhibited by 93.75% at 375 $\mu\textrm{g}$/mL. Hexane fraction of ixeris dentate exhibited 68.56% inhibition against MCF-7 human breast adenocarcinoma cells at 500 $\mu\textrm{g}$/mL. Hexane fraction of Ixeris dentata exhibited 84.91% inhibition against Hep 3B human hepatocellular carcinoma cells at 500 $\mu\textrm{g}$/mL. From these results, it is considered that Ixeris dentata has strong antimutagenic and anticancer effects in vitro. However, these extracts and fractions did not show any cytotoxic effect against 293 cells.

Fracture resistance of zirconia and resin nano ceramic implant abutments according to thickness after thermocycling (지르코니아와 레진나노세라믹 임플란트 지대주의 두께에 따른 열순환 후 파절저항)

  • Lee, Jung-Won;Cha, Hyun-Suk;Lee, Joo-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.55 no.2
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    • pp.144-150
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    • 2017
  • Purpose: The aim of this in vitro study is to investigate load bearing capacity of esthetic abutments according to the type of material and wall thickness. Materials and methods: 70 specimens equally divided into seven groups according to their abutment wall thicknesses. The abutments prepared with titanium 0.5 mm wall thickness were used as a control group (Ti-0.5), whereas zirconia abutments and resin nano ceramic abutments with wall thickness 0.5 mm, 0.8 mm and 1.0 mm were prepared as test groups (Zir-0.5, Zir-0.8, Zir-1.0 and RNC-0.5, RNC-0.8, RNC-1.0). All specimens were tested in a universal testing machine to evaluate their resistance to fracture and all of them underwent thermo-cycling before loading test. Mean fracture values of the groups were measured and statistical analyses were made using two-way ANOVA. Results: Zir-1.0 showed the highest mean strength ($2,476.3{\pm}342.0N$) and Zir-0.8 ($1,518{\pm}347.9N$), Ti-0.5 ($1,041.8{\pm}237.2N$), Zir-0.5 ($631.4{\pm}149.0N$) were followed. The strengths of RNC groups were significantly lower compared to other two materials (RNC-1.0 $427.5{\pm}72.1$, RNC-0.8 $297.9{\pm}41.2$) and the strengths of all the test groups decreased as the thickness decreases (P < .01). RNC-0.5 ($127.4{\pm}35.3N$) abutments were weaker than all other groups (P < .05). Conclusion: All tested zirconia abutments have the potential to withstand the physiologic occlusal forces in anterior and posterior regions. In resin nano ceramic abutments, wall thickness more than 0.8 mm showed the possibility of withstanding the occlusal forces in anterior region.

Induction of Somatic Hybrid by Protoplast Fusion between Populus koreana × P. nigra var. italica and P. euramericana cv. Guardi (수원포플러와 구아디 포플러 원형질체(原形質體) 융합(融合)에 의한 체세포잡종체(體細胞雜種體) 유도(誘導))

  • Park, Young Goo;Kim, Jung Hee;Son, Sung Ho
    • Journal of Korean Society of Forest Science
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    • v.81 no.3
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    • pp.273-279
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    • 1992
  • Protoplasts isolated from leaf mesophyll tissues of Populus koreana ${\times}$ P. nigra var. italica were fused with those of P. euramericana cv. Guardi. Well expended healthy leaves of 5 to 7 week-old-plantlet grown in vitro were used as source materials. Leaves from P. koreana ${\times}$ P. nigra var. italica and P. euramericana cv. Guardi were digested in enzyme solution I (2.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v) and enzyme solution II (1.0% Cellulase, 1.2% Hemicellulase, 0.4% Macrozyme, 2.0% Driselase, 0.05% Pectolyase ; w/v), respectively, The highest frequency of fusion among the protoplasts originated from the two source materials was approximately 21% using 40% PEG or 15% dextran. In addition, fusion frequency was enhanced by incorporating 30mM of $Ca^{2+}$ in eluting solution at pH 10.5. Dividing cells and/or mint-calli were obtained by culturing the fusion products in a liquid 8p-KM medium supplemented with 0.6M sucrose, $0.45{\mu}M$ 2, 4-D, and $0.5{\mu}M$ BA. Shoots were regenerated from the fusion product-derived calli after culture on MS medium containing $5.0{\mu}M$ zeatin. To verify the putative hybrid or cybrid, SDS-PAGE was carried out. From the 24 regenerants, just two plants showed intermediate protein band patterns compared with those of the original source plants.

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Comparison of Agronomic Characteristics, Forage Production and Quality of Kenaf (Hongma 300), Maize (Kwangpyeongok) and Sorghum × Sudangrass Hybrids (Jumbo) in Middle Region of Korea (중부지역에서 케나프(홍마 300), 옥수수(광평옥) 및 수수×수단그라스 교잡종(점보)의 생육특성, 수량성 및 품질비교)

  • Hwang, Tae-Young;Ji, Hee Chung;Kim, Ki Yong;Lee, Sang-Hoon;Lee, Ki-Won;Choi, Gi Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.2
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    • pp.152-158
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    • 2015
  • This study was conducted from 2013 to 2014 in the middle region of Korea to compare the agronomic characteristics, forage production, and quality of kenaf, maize, and a sorghum ${\times}$ sudangrass hybrid. The three crops used in this study were kenaf "Hongma 300," maize "Kwangpyeongok," and the sorghum ${\times}$ sudangrass hybrid "Jumbo" (headless type). The sugar contents of the kenaf, maize, and sorghum ${\times}$ sudangrass hybrid were 5.4%, 4.4%, and 2.5%, respectively. The emergence rates and lodgings of the three crops were sound, as they were more than 80% and 1.5, respectively. The fresh yields of kenaf, maize, and the sorghum ${\times}$ sudangrass hybrid were 117,521 kg/ha, 73,336 kg/ha, and 133,334 kg/ha, respectively, and the dry matter (DM) yields were 28,598 kg/ha, 19,951 kg/ha and 20,083 kg/ha, respectively. The DM yield of kenaf was significantly the highest among the three crops (p<0.05). However, the total digestible nutrients (TDN) contents of the kenaf, corn, and sorghum ${\times}$ sudangrass hybrid were 43.3 %, 72.0 %, and 54.8 %, respectively, and the in vitro digestible dry matter (IVDDM) yields were 44.4%, 73.7%, and 59.6 %, respectively; therefore, the TDN yield of the corn was 13 % significantly higher than that of the kenaf (p<0.05). The TDN yields of the kenaf, corn, and sorghum ${\times}$ sudangrass hybrid were 12,472 kg/ha, 14,350 kg/ha, and 11,001 kg/ha, respectively. Among the three crops, the content of neutral detergent fiber (NDF) and acid detergent fiber (ADF) were highest in the kenaf. This study shows that the kenaf had a good forage productivity but a low forage quality.

High frequency somatic embryogenesis through leaf explant-derived callus culture in Muscari armeniacum cv. 'Early Giant' (무스카리 'Early Giant' 잎 절편 유래 캘러스 배양을 통한 고빈도 체세포배 발생)

  • Lee, Hyang-Bun;Jeon, Su-Min;Chung, Mi-Young;Han, Jeung-Sul;Kim, Chang-Kil;Lim, Ki-Byung;Chung, Jae-Dong
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.69-74
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    • 2012
  • Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.

Delayed closure effect in preterm infants with patent ductus arteriosus (미숙아 동맥관개존증의 지연된 폐쇄가 예후에 미치는 영향)

  • Lee, Hyun Ju;Sim, Gyu Hong;Jung, Kyung Eun;Lee, Jin A;Choi, Chang Won;Kim, Ee Kyung;Kim, Han Suk;Kim, Beyong Il;Choi, Jung-Hwan
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1065-1070
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    • 2008
  • Purpose : This study aims to determine whether early closure (within 7 d) of significant patent ductus arteriosus (PDA) with indomethacin or ligation reduces neonatal morbidity when compared with delayed closure (after 7 d). Methods : Fifty-eight extremely-low-birth-weight infants admitted to the NICU of Seoul National University Hospital from April 2005 to May 2007 with PDA were studied retrospectively. Results : The mean gestational age (GA) was $26{\pm}2weeks$ (range, 23-32 wk), and the birth weight was $782{\pm}146g$ (range, 430-990 g). The delayed closure group was associated with early GA ($25.7{\pm}1.7wk$ vs $27.1{\pm}2.0wk$, P=0.013), in vitro fertilization (IVF) (55% vs 24%, P=0.017), and the absence of preeclampsia (5% vs. 34%, P=0.013). There was no difference in ductal size between the early closure and delayed closure groups. The incidence of bronchopulmonary dysplasia (95% vs 65%, P=0.012) and intraventricular hemorrhage (70% vs. 39%, P=0.027) increased in the delayed closure group. Using regression analysis adjusted for gestational age, delayed closure correlated positively with the duration of ventilator support (P=0.008), hospitalization (P=0.020), time to full enteral feeding (P<0.001), and total parenteral nutrition (P=0.010). Conclusion : Delayed closure of the hemodynamically significant patent ductus arteriosus in extremely-low-birth-weight infants is significantly related to the development of various morbidities. Thus, early closure of PDA is needed within the first week of life.

Effect of Maturity at Harvest and Wrap Colors on the Quality of Round Baled Rye Silage (수확시 숙기 및 비닐색이 호밀 라운드베일 사일리지 품질에 미치는 영향)

  • Kim, J.G.;Kim, D.A.;Chung, E.S.;Seo, S.;Kim, J.D.;Ham, J.S.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.4
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    • pp.355-362
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    • 1999
  • This experiment was conducted to evaluated the effect of maturity at harvest and wrap colors on the quality of round baled rye(Secal cereale L.) silage at the forage experimental field, Grassland and Forage Crops Division, National Livestock Research Institute, Suwon in 1998. The experiment was consist of split-plot design with three repications. The main plots were three different harvesting stages : boot, heading, and flowering stages, the subplots were wrap colors : white, black, and light green color. Acid detergent fiber(ADF) and neutral detergent fiber(NDF) of rye silage were increased with delayed harvesting date, but there were no significant differdence among wrap colors. However, average in vitro dry matter digestibility (IVDMD) of rye silage with white color wrap tended to have higher than those of other colors. The silage pH of heading stage was highest (5.12), and that of flowering stage was lowest (4.57) in different harvest stages. Among tested wrap colors, white color wrap resulted on lowest pH than others (p<0.05), but there were no significant difference between black and light green color. Dry matter (DM) content of rye silage were increased as harvest stage progressed (p<0.05). However, DM content of white color was highest among warp colors, but there was signifiant difference (p<0.05). Ammonia-N content of silage harvested at boot stage was the highest in harvest stage, but there was no difference between heading and flowering stages. Among wrap colors tested, ammonia-N content of black color was highest, but there was no significant difference. The number of lactic acid bacteria of white colors was highest in different colors, and that of flowering stage was highest among harvest stage. Acetic and butyric acid contents were decreased as harvest stage progressed, and lactic acid was increased from 6.33 to 7.98%. However, wrap colors did not influence lactic acid concentration. Among different wrap colors, outside temperature of rye silage was affected by air temperature, but effect of inside temperature was minimal. Black color wrap increased inside temperature by $3{\sim}5^{\circ}C$. The results of this study indicate that rye should be harvested for round bale silage after heading stage. The quality of rye silage wrapped white and light green color will increase slightly.

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