• Title/Summary/Keyword: 계대 배양

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Developmental Potentials of Clone Embryos Derived from Bovine Fetal Fibroblast Cells (소 태아섬유아세포 유래 복제란의 발육능에 관한 연구)

  • Cheong, H.T.;Kwon, D.J.;Park, Y.S.;Hwang, H.S.;Park, C.K.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.49-57
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    • 2000
  • This study was conducted to investigate the developmental potential of cloned embryos derived from bovine fetal fibroblast cells, and the effect of quiescent treatment, passage number and origin of donor cells on in vitro development of cloned embryos. Fetal skin and liver-derived fibroblast cells were transferred to enucleated oocytes after serum starvation or nontreatment (cycling). After electrofusion. reconstituted embryos were activated with $Ca^{++}$-ionophore and cycloheximide, and cocultured for 7~9 days with BRL cells. Some blastocysts were transferred to recipient cows 7~8 days post estrus. The development rate to the blastocyst stage of serum starved cell-derived embryos was higher (25.3%) than that of actively dividing cells-derived embryos (15.9%), The rates of blastocyst formation were 23.1~25.0% after transfer of cell passaged 4 to 6 times, and 23.8 and 25.2% after transfer of fetal skin and liver cells, respectively. After embryo transfer, 34.4% and 15.6% of recipient cows were pregnant on Day 60 and 120, respectively, and one male calf was produced from skin-derived vitrified blastocyst. The result of this study showed that the development of cloned embryos. was enhanced by quiescent treatment, but did not different among the cells passaged 4 to 6 times, and between skin and liver cells. This result also confirms that offspring can be obtained from the vitrified clone embryo derived from fetal skin cell.

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Transformation of Adult Mesenchymal Stem Cells into Cardiomyocytes with 5-azacytidine: Isolated from the Adipose Tissues of Rat (성체 백서의 지방조직에서 추출한 중간엽 줄기세포의 5-azacytidine을 이용한 심근세포 분화 유도)

  • Choe Ju-Won;Kim Yong-In;Oh Tae-Yun;Cho Dai-Yoon;Sohn Dong-Suep;Lee Tae-Jin
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.511-519
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    • 2006
  • Background: Loss of cardiomyocytes in the myocardial infarction leads to regional contractile dysfunction, and necrotized cardiomyocytes in infracted ventricular tissues are progressively replaced by fibroblasts forming scar tissue. Although cardiomyoplasty, or implantation of ventricular assist device or artificial heart was tried in refractory heart failure, the cardiac transplantation was the only therapeutic modality because these other therapeutic strategies were not permanent. Cell transplantation is tried instead of cardiac transplantation, especially bone marrow is the most popular donated organ. But because bone marrow aspiration procedure is invasive and painful, and it had the fewer amounts of cellular population, the adipose tissue is recommended for harvesting of mesenchymal stem cells. Material and Method: After adipose tissues were extracted from abdominal subcutaneous adipose tissue and intra-abdominal adipose tissue individually, the cellular components were obtained by same method. These cellular components were tried to transformation with the various titers of 5-azacytidine to descript the appropriate concentration of 5-azacytidine and possibility of transformation ability of adipose tissue. Group 1 is abdominal subcutaneous adipose tissue and Group 2 is intra-abdominal adipose tissue-retroperitoneal adipose tissue and omentum. Cellular components were extracted by collagenase and $NH_4Cl$ et al, and these components were cultured by non-induction media - DMEM media containing 10% FBS and inducted by none, $3{\mu}mol/L,\;6{\mu}mol/L,\;and\;9{\mu}mol/L$ 5-azacytidine after the 1st and 2nd subculture. After 4 weeks incubation, tile cell blocks were made, immunostaining was done with the antibodies of CD34, heavy myosin chain, troponin T, and SMA. Result: Immunostaining of the transformed cells for troponin T was positive in the $6{\mu}mol/L\;&\;9{\mu}mol/L$ 5-azacytidine of Group 1 & 2, but CD34 and heavy myosin chain antibodies were negative and SMA antibody was positive in the $3{\mu}mol/L\;&\;6{\mu}mol/L$ 5-azacytidne of Group 2. Conclusion: These observations confirm that adult mesenchymal stem cells isolated from the abdominal subcutaneous adipose tissues and intra-abdominal adipose tissues can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair.

Studies on the Associative N2-fixing Microbes in Crop Root Systems -I. Associative Nitrogen Fixation by Azospirillum Isolated from Weed Roots grown Rice Paddy Field and its Distribution (작물근계(作物根系) 협생질소(協生窒素) 고정미생물(固定微生物)에 관(關)한 연구(硏究) -I보(報) : 논 토양(土壤) 잡초근계(雜草根系) 협생질소(協生窒素) 고정미생물(固定微生物)의 분포(分布)와 질소고정(窒素固定))

  • Ryu, Jin-Chang;Lee, Sang-Kyu;Park, Jun-Kyu;Ko, Jae-Young;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.17 no.4
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    • pp.415-427
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    • 1984
  • This experiment was carried out to investigate the distribution of associative nitrogen fixers, Azospirillum spp. and their nitrogenase activities measured by ARA in the rhizosphers of rice, soybean, and weed grown in the rice paddy field at ear formation stage of rice. Nitrogenase activities produced by Azospirillum spp. enriched from histophere ranged from 16 to 53 n mole/tube/hr.. High nitrogen fixing activities more than 30 n mole/tube/hr. were observed in the histophere of the Echinochloa crus-galli L., Finbristylis miliacea L., and Monochoria vaginalis var.. Nitrogen fixing activities of Azospirillum spp. obtained from single colonies which originated from the rhizoplane of rice (pot-kwang var.), Finbris tylis miliacea L., Monochoria vaginaliz var., Glycine max L. were higher over 100 n mole/tube/hr. than those histophere. Genus of Azopsirillum isoltated from roots of the Graminease (Oriza sativa L., $C_3$-plant, Echinochloa crus=galli L., $C_4$-plant, Cyperus difforuis L.. $C_4$-plant), and Aeschynomene indica L. (Leguminosae, $C_3$-plant) was identified as A. brasilense. However, both strains, A. lipoferum and A. brasilense ($nir^-$ or $nir^+$ strain) were isolated from other plant roots, Both $nir^-$ and $nir^+$ strains of A. brasilense were associated with the same host plant.

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Nucleotide Sequence Analysis of the Hemagglutinin-Neuraminidase Gene of Urabe AM-9 Strain (Urabe AM-9 볼거리 백신주의 Hemagglutinin-Neuraminidase 유전자 염기서열 분석)

  • Lee, Joo Yeon;Kim, Jee Hee;Lee, Jin Soo;Park, Ji Ho;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.83-93
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    • 2000
  • Purpose : Urabe AM-9 strain was known to be associated with increased aseptic meningitis. The reason for high incidence of vaccine-associated meningitis was known that nucleotide(nt) substituted form G to A at position 1081 of the hemagglutinin-neuraminidase(HN) gene and therefore, glutamic acid changed to lysine at amino acid 335. We assessed by comparing nt sequence of the HN gene form Urabe AM-9 strain with wild strain and documented the correlation between nt substitution and vaccine-associated meningitis. Methods : Two lots of Urabe AM-9 vaccine distributed in Korea and mumps wild strains isolated from 1998 through 1999 were analysed. Analysis was made by nt sequencing following amplification of HN gene by RT-PCR. Results : Nucleotide substitution at position 343, 1476, 1570 was not found in both Urabe AM-9 vaccines and wild strains. But analysis of vaccine strains and wild strains isolated from patients revealed substitution from G to A at nt 1081 of the HN gene. Therefore, it encodes lysine instead of glutamic acid at amino acid 335. There was no mixture from of G and A at nt 1081. Nt at 1470 of one lot of Urabe AM-9 vaccines changed from C to A after Vero cell passage. Nt at 1727 of vaccines and wild strains was substituted A to G, so it encodes glycine instead of aspartic acid. Conclusion : Nucleotide analysis of HN gene revealed that nt 1081 of Urabe AM-9 vaccines and wild strains had wild type AAA($Lys^{335}$) instead of variant type GAA($Glu^{335}$). The results of this study suggest that there was a probability of vaccine-associated meningitis due to Urabe AM-9 in Korea before. But incidence of actual side effect was not evaluated because there was no reporting system in Korea.

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Drug resistance of Shigella and Salmonella and the inhibition and elimination of drug resistance (이질균(痢疾菌) 및 살모내라의 약제내성(藥劑耐性), 내성화방지(耐性化防止) 및 제거(除去))

  • Chun, Do-Ki;Seol, Sung-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.14 no.1
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    • pp.27-37
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    • 1979
  • Ninety-five strains of Shigella, 70 of Salmonella paratyphi A, and 230 of Salmonella typhi were tested for their resistance to drugs. Also studied was the inhibition and elimination of drug resistance. All except one strain of Shigella consisted of 79 Sh. flexneri and 16 Sh. sonnei were multiply resistant to chloramphenicol, tetracycline, streptomycin, and splfisomidine. Among them, 70 strains were resistant to ampicillin and carbenicillin, 80 to trimethoprim-sulfamethoxazole, 22 to nalidixic acid, and one to kanamycin, but strain resistant to gentamicin, cephaloridine, and rifampin was not encountered. All strains of S. paratyphi A and S. typhi were susceptible to drugs tested, except sulfisomidine and rifampin, for which all S. paratyphi A were slightly resistant to sulfisomidine and the majority of S. paratyphi A and S. typhi were slightly resistant to rifampin. Approximately 80% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation, and the resistance was considered to be mediated by R plasmids. The frequency of transfer of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to the recipients. Drug-resistant Shigella strains successively subcultured in nutrient agar stabs contained clones resistant to drugs and those susceptible to drugs, but the ratio of resistant and susceptible clones varied by strains. The multiply drug-resistant S. typhi and Shigella strains were found to not lose completely their drug resistance by subculture in media. Acriflavine has some effect on the elimination of drug resistance mediated by R plasmids, but the effect varied markedly by strains. Atabrine has no effect among strains tested. The combination of drugs increased the drug actions in majority of cases with synergistic or additive effects.

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In Vitro Plant Regeneration for Mass Propagation of Epimedium koreanum Nakai (삼지구엽초의 다량번식을 위한 기내 식물체 분화)

  • Han, Young-Hee;Choi, Byoung-Ryourl;Soh, Ho-Seob;Lee, Seong-Jae;Choi, Young-Jin;Kim, Se-Young
    • 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.

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A Study on the Screening of the Novel Genes Associated with Lysosomal Trafficking and Mutation Detection in Fibroblasts of the Patients with Mucolipidosis type II and III (리소좀 교통 이상을 초래하는 뮤코지방증 2형과 3형 환자의 섬유아세포를 이용한 신규 유전자 탐색 및 돌연변이에 대한 연구)

  • Song, Seng Mi;Chang, Soo Hee;Paik, Kyung Hoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.5 no.1
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    • pp.65-75
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    • 2005
  • Purpose: To understand genetic differences and similarities between mucolipidosis and control. Methods: Using the fibroblast of the mucolipidosis II and control, forward and reverse subtracted libraries were constructed. Among these clones, we investigated mutations in the GNPTA (MGC4170) gene, which codes for the ${\alpha}/{\beta}$ subunits of phosphotransferase, and in the GNPTAG gene, which codes for the ${\gamma}$ subunits in 5 Korean patients with mucolipidosis type II or IIIA. Result: Several differentially expressed cDNAs were cloned and their sequences were determined. Mutation analysis of the interested gene, GNPTA was performed and we identified 7 mutations in the GNPTA gene, but none in the GNPTAG gene. The mutations in type II patients included p.Q104X(c.310C>T), p.R1189X(c.3565C>T), p.S1058X(c.3173C>G), p.W894X(c.2681G>A) and p.H1158fsX15(c.3474_3475delTA), all of which are non-sense or frame shift mutations. However, a splicing site mutation, IVS13+1G>A (c.2715+1G>A) was detected along with a non-sense or a frame shift mutation (p.R1189X or p.E858fsX3(c.2574_2575delGA)) in two mucolipidosis type IIIA patients. Conclusion: This report shows that mutations in the GNPTA gene coding for the ${\alpha}{\beta}$subunits of phosphotransferase, and not mutations in the GNPTAG gene, account for most of mutations found in Korean patients with mucolipidosis type II or IIIA.

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A Study on the Effect of Metals on Bacteria Adhesion to Zeolite as Bio-media Materials (제올라이트를 이용한 생물막 형성시 미생물의 부착에 금속이 미치는 영향에 관한 연구)

  • Kim, Jae Keun;Park, In Sun;Park, Jae-Woo
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.29 no.3B
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    • pp.303-310
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    • 2009
  • Natural zeolite is widely used as sorbents and bio-media materials because it is cheap as well as it has efficient porous structures and large cation exchange. In this study, the effect of metal cations $(Na^+,\;Ca^{2+},\;Mg^{2+},\;Al^{3+})$ adsorbed to natural zeolite on the microorganism attachment was investigated. Metal-modified zeolites (MMZ) were prepared with 0.01 M, 0.02 M and 0.1 M NaCl, $CaCl_2$, $MgCl_2$ and $AlCl_3$ solutions respectively, which concentrations were equivalent to 10%, 20% and 100% of cation exchange capacity (CEC) of natural zeolite. Pseudomonas putida was used as microorganism which was cultivated in Beef Extract Medium at $26^{\circ}C$. The microorganism attachment to MMZ was increased more than natural zeolite. The amount of bacterial adhesion to MMZ and natural zeolite were $Mg^{2+}>natural>Na^+>Al^{3+}>Ca^{2+}$ under 10% of CEC, $Mg^{2+}>Ca^{2+}>Al^{3+}>natural>Na^+$ under 20% of CEC and $Ca^{2+}>Mg^{2+}>natural>Al^{3+}>Na^+$ under 100% of CEC. Especially, Mg-modified zeolite (Mg-MZ) showed the highest amount of bacterial adhesion, which increased the microorganism attachment 60% higher than natural zeolite under 10% of CEC. However, the amount of bacterial adhesion was decreased as the concentration of metal cations modified to zeolite were increased, showing that the increased amounts were 60% under 10% of CEC, 50% under 20% of CEC and 10% under 100% of CEC in Mg-MZ. Additionally, the effect of $Mg^{2+}$ in solution on the bacterial adhesion was investigated in order to compare it with the effect of $Mg^{2+}$ adsorbed to zeolite. The maximum quantity of bacterial adhesion to Mg-MZ was not different from the amount of microorganism attachment to the natural zeolite when $Mg^{2+}$ solution was added.