• 제목/요약/키워드: Cell isolation

검색결과 1,079건 처리시간 0.068초

Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library

  • Kim, Min-Soo;Choi, Hye-Young;Choi, Yong-Soo;Kim, Jhin-Gook;Kim, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.993-1001
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    • 2007
  • The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.

Naphthalene 분해균주 Alcaligenes sp. A111의 분리 및 특성 (Isolation and Characterization of a Naphthalene-Degrading Strain,Alcaligenes sp,A111)

  • 오희목;강정현;이창호;박찬선;안성구;윤병대;고영희
    • 한국미생물·생명공학회지
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    • 제22권4호
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    • pp.423-429
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    • 1994
  • A bacterial strain which formed a distinct colony on agar plate containing naphthalene as a vapor phase and grew well ina liquid minimal medium was isolated and identified as Alcaligenes sp. A111. Optimum temperature and pH for the cultivation of Alcaligenes sp. A111 were 30$\cir$C and 7.0, respectively. Cell growth increased dramatically from 12 hours after inoculation and revealed a stationary phase at about 48 hours. Relative growth rate ($\mu$')increased hyperbolically depending on the conceration of naphthalene up to 500 ppm and reached to the maximum value pf 2.8/day, but $\mu$' didn't change within a range of 500~4000 ppm naphthalene. NH$_{4}$Cl or NH$_{4}$NO$_{3}$ was preferrd as a nitrogen source and a P : N ratio by weight og 6 : 1 was favorable to cell growth. Alcaligenes sp. A111 utilized the intermediates of degradation of naphthalene and showed tolerance to benzene, toluene, and octane. therefore, it is suggested that Alcaligenes sp. A111 could be effectively used for the biological treatment of wastewater containing naphthalene in the presence of some aromatic compounds.

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Spontaneous Electrical Activity in Cerebellar Purkinje Neurons of Postnatal Rats

  • Nam, Sang-Chae
    • The Korean Journal of Physiology and Pharmacology
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    • 제1권4호
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    • pp.355-366
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    • 1997
  • Although cerebellar Purkinje cells display spontaneous electrical activity in vivo and in slice experiments, the mechanism of the spontaneous activity generation has not been clearly understood. The aim of this study was to investigate whether cerebellar Purkinje cells of postnatal rats generate spontaneous electrical activity without synaptic inputs. Dissociated cerebellar Purkinje cells were used for reducing synaptic inputs in the present study. Cerebellar Purkinje cells with dendrites were dissociated from postnatal rats using enzymatic treatment followed by mechanical trituration. Spontaneous electrical activities were recorded from dissociated cells without any stimulus using whole-cell patch clamp configuration. Two types, spontaneously firing or quiescent, of dissociated Purkinje cells were observed in postnatal rats. Both types of cells were identified as Purkinje cells using immunocytochemical staining technique with anti-calbindin after recording. Spontaneously active cells displayed two patterns of firing, repetitive and burst firings. Two thirds of dissociated Purkinje cells displayed repetitive firing and the rest of them did burst firing under same recording condition. Repetitive firing activities were maintained even after further isolation using either physical or pharmacological techniques. Neither high magnessium solution nor excitatory synaptic blockers, AP-5 and DNQX, block the spontaneous activity. These results demonstrate that spontaneous electrical activity of isolated cerebellar Purkinje cells in postnatal rats is generated by intrinsic membrane properties rather than synaptic inputs.

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Protoplast Isolation and Regeneration of Fertile Plants from Arabidopsis Trp Mutant, trp1-100

  • Lim, Seon-hee;Kim, Young-soon;Lee, Eui-seung;Rose, Alan;Last, Robert;Cheong, Hyeon-sook
    • Animal cells and systems
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    • 제2권2호
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    • pp.239-242
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    • 1998
  • Arabidopsis trp1 mutant plants, deficient in phosphoribosyI anthranilate transferase (PAT) activity, accumulate anthranilate compounds, which render them blue fluorescence. The visible phenotype of trp1 makes the PAT gene an excellent reporter gene in the mutant. In order to develop a system for the homologous recombination using the phenotypic characteristic of trp1-100, we established optimum conditions for the isolation and regenera tion of protoplast from auxin-conditioned, trp1-100 root cultures. Trvptophan had to be supplemented in the germination medium for the efficient cell division and subsequent plant regeneration. When 10 uM tryptophan was added to the germination medium, we obtained the highest yield of protoplasts ($3{\times}10^6 cells/g$) and the best viability (92%). Thirty percent of root protoplast derived from meristematic cells underwent cell division within 5 days in callus-induction medium. Regenerated rosette leaves (2-3 mm) were transferred to rooting medium and finally acclimated to the soil for flowering.

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독립된 접지면을 갖는 EBG 구조를 이용한 이중 대역 마이크로스트립 패치 안테나 사이의 격리도 향상 (Isolation Enhancement between Two Dual-Band Microstrip Patch Antennas Using EBG Structure without Common Ground Plane)

  • 최원상;이홍민
    • 한국전자파학회논문지
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    • 제23권3호
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    • pp.306-313
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    • 2012
  • 본 논문에서는 UMTS Tx 대역(1.92~1.98 GHz)과 Rx 대역(2.11~2.17 GHz)에서 동작하는 이중 대역 E 슬롯 마이크로스트립 패치 안테나 사이의 격리도 향상을 위한 EBG 구조를 제안하였다. 제안된 EBG 구조는 이중 대역에서 두 안테나 사이의 격리도를 향상시키기 위하여 서로 다른 크기의 변형된 버섯 모양(mushroom-type) 구조로 단위 셀들을 배열하였다. 제안된 구조는 패치 안테나와 접지 면을 공유하지 않으며, 설계되어진 단위 셀의 크기는 각각 $15.6mm{\times}4mm$, $17.4mm{\times}4mm$이다. 제안된 안테나의 전체 크기는 $210.5mm{\times}117mm$이고, FR-4(비유전율=4.6, 기판 두께= 3.93 mm) 기판 상에 설계되어졌다. 제작된 안테나의 측정 결과, 제안된 EBG 구조의 사용으로 UMTS Tx 대역과 Rx 대역에서 각각 9 dB, 12 dB의 격리도 향상을 나타내었다.

Validation of the production quality and therapeutic efficacy of 47Sc through its anti-cancer effects against EGFR-targeted non-small cell lung cancer

  • Da-Mi Kim;So-Young Lee;Jae-Cheong Lim;Eun-Ha Cho;Ul-Jae Park
    • 대한방사성의약품학회지
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    • 제8권1호
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    • pp.9-15
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    • 2022
  • Anti-cancer and therapeutic effects using therapeutic radioisotopes have been demonstrated by various studies, and it is well-known that therapeutic radioisotopes are useful in cancer treatment. Recently, one of the therapeutic radioisotopes, scandium is emerging as a radioisotope applicable to PET imaging (43Sc, 44Sc) and therapy (47Sc) in cancer theranostic approach. However, 47Sc has little known radiobiological and therapeutic efficacy compared to other therapeutic radioisotopes. Here, we investigated the quality and therapeutic efficacy of 47Sc radioisotope produced by our production/isolation technology at the research reactor 'HANARO' in KAERI (Korea Atomic Energy Research Institute). We showed that the therapeutic efficacy of 47Sc, produced by our production/isolation technology, effectively suppressed epidermal growth factor receptor (EGFR)-targeted non-small cell lung cancer (NSCLC) cells. Consequently, these results suggest that the high quality of the produced 47Sc by our production/isolation technology enables the development of therapeutic strategies for cancer treatment and radiopharmaceuticals using 47Sc.

Isolation and characterization of feline endometrial mesenchymal stem cells

  • Mi-Kyung Park;Kun-Ho Song
    • Journal of Veterinary Science
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    • 제25권2호
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    • pp.31.1-31.8
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    • 2024
  • Background: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. Objectives: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. Methods: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. Results: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 -, CD34 -, CD45 -, CD9+, and CD44+. Conclusions: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

Construction of Astaxanthin Overproducing Strain of Phaffia rhodozyma by Protoplast Fusion

  • Koh, Moo-Suk;Kim, Sang-Moon
    • Journal of Microbiology and Biotechnology
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    • 제2권1호
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    • pp.46-49
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    • 1992
  • The availability of Phaffia rhodozyma as an astaxanthin sources in the aquaculture industry is limited because of the low carotenoid content of natural isolate. In this study, we have used the protoplast fusion technique to construct cell hybrids with an increased content of astaxanthin from P. rhodozyma. Cell hybrids (F307 and F406) obtained were very stable and produced considerably more astaxanthin (> 1 mg/g yeast) than the wild parent. Karyogamy was confirmed by the isolation of recombinants after mitotic segregation of parental auxotrophic genetic markers, the increased amount of chromosomal DNA/cell and the presence of single nucleus/cell.

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암모니아산화세균 Brevundimonas diminuta의 분리 및 암모니아 산화 특성 (Isolation and Characteristics of Novel Ammonia Oxidizing Bacteria Brevundimonas diminuta)

  • 권혁구;정준오
    • 한국환경보건학회지
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    • 제33권4호
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    • pp.293-298
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    • 2007
  • The microorganism for ammonia gas removal was isolated from composting product. This was identified as Brevundimonas diminuta by morphological, biochemical characteristics study and 16S rDNA sequence analysis. Optimal incubation temperature for cell growth and oxidizing ability of $NH_4-N$ was $30^{\circ}C$ and optimal initial pH was 7. Glucose affected the growth of cell and the removal of $NH4^+$. The growth rate of the isolates were increased when grown in the presence of 0.05-1%(w/v) glucose in the selective medium and lurker increases in glucose concentration to 2% caused significant decreases in the cell growth and oxidizing ability of $NH4^+$.

A Quinolone Alkaloid, from the Aleurone Layer of Oryza sativa cv. Mihyangbyo, Inhibits Growth of Cultured Human Leukemia Cell

  • Chung, Ha-Sook
    • Preventive Nutrition and Food Science
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    • 제7권2호
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    • pp.119-122
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    • 2002
  • Oryza sativa cv. Mihyangbyo is one of several recently developed varieties of rice; characterized by high levels of aromatic components, which may increase its sensory and nutritional properties. In conjunction with our continuing investigation of bioactive components of improved grain varieties, a quinolone alkaloid was isolated from the n-butanol soluble fraction of the aleurone layer of Oryza sativa cv. Mihyangbyo (Gramineae) through activity-guided fractionation and isolation. The compound exhibited moderate antineoplastic activity in a human leukemia cell line (U937) with an $IC_{50}$/ value of 118.1 ug/mL, based on the MTT(3-[4, 5]dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) cell proliferation assay. The chemical structure of the functional compound was determined, based on physical and spectroscopic characteristics.