• 제목/요약/키워드: Cell-based assay

검색결과 663건 처리시간 0.028초

단핵구 분화에 대한 Fibronectin 및 그 단편의 역할 (A Possible Role of Fibronectin on the Differentiation of Monocyte to Macrophase)

  • Ok Sun Bang;You
    • 한국동물학회지
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    • 제36권4호
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    • pp.514-521
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    • 1993
  • Monocyte interaction with fibronectin (FN) mediates specific cell surface receptors and results in cell attachment and differentiation. Several cell-mediated activities for various fragments of FN have been documented. To investigate the regulatory mechanisms of monocyte differentiation by cell binding domains of FN and their receptors, cell attachment-, cell migration-, and its respective inhibition assay were carried out. Monocyte recognizes 38-kDa domain distinctively from its recognition of 85-kDa domain, and the heparin-binding site of the 38-kDa fragment is not involved in monocyte adhesion. Based on these experimental results, it can be suggested that monocvte/macrophase interacts with at least two different sites in FN, which is critical step in cell adhesion and (or) migration.

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아스코르브산의 3차원 줄기세포 배양체에 대한 증식 및 분화 효과 분석 (The effects of ascorbic acid on the morphology and cell proliferation of three-dimensional stem cell spheroids)

  • 이현진;여성일;박준범
    • 대한치과의사협회지
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    • 제55권9호
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    • pp.592-603
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    • 2017
  • Purpose: The effects of various concentrations of ascorbic acid on stem cell spheroids derived from intraoral areas are not known yet. Thus, the purpose of this study is to evaluate the effects of different concentrations of ascorbic acid on the morphology and cellular viability of stem cell spheroids derived from the gingival tissues. Materials and Methods: Stem cells were plated onto silicon elastomer-based concave microwells and grown in the presence of ascorbic acid at concentrations ranging from 0.003% to 0.3%. The morphology of the cells was viewed under an inverted microscope at day 1, 2, 3 and 5. Qualitative live/dead assay and quantitative cellular viability using Cell Counting Kit-8 were performed on day 2 and day 5. Results: Gingiva-derived stem cells formed spheroids irrespective of ascorbic acid concentration in silicon elastomer-based concave microwells. Increase in the diameter of spheroid were seen with higher concentrations of ascorbic acid. Higher cellular viability was seen in higher concentrations of ascorbic acid. Conclusion: Within the experimental setting, the application of ascorbic acid on stem-cell spheroids produced an increase in the size and higher viability with higher dosage. It can be suggested ascorbic acid be applied with stem cell spheroids for tissue engineering purposes.

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Phorbol 12-myristate 13-acetate (PMA) 처리로 유도되는 THP-1 세포의 초기 부착에 관한 다양한 인자의 효과 (Effect of Various Factors on Early THP-1 Cell Adhesion Induced Phorbol 12-Myristate 13-Acetate (PMA))

  • 조용삼;신지현;최태생
    • 생명과학회지
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    • 제18권7호
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    • pp.952-957
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    • 2008
  • 본 실험에서는 THP-1 세포의 PMA에 의하여 유도되는 초기 세포부착에 관한 메카니즘을 이해하기 위하여 다양한 요인(혈청, 신규 단백질의 합성, 세포 골격 저해제, 단백질 인신화 저해제)들의 효과를 조사하였다. 또한 본 실험에서는 이들 세포부착의 정도를 일반적으로 세포증식 분석에 사용되고 있는 SRB염색법을 도입하여 세포부착 분석에 간편한 방법의 조건을 확립하였다. PMA에 의한 초기 세포부착에는 배양액중의 혈청의 유무는 영향이 없었으나, 신규 단백질의 합성이 요구되는 것을 확인하였다. 또한 이들 초기 세포부착에 PMA처리에 의한 PKC의 활성화는 필수적이나, 그 하류 활성화 인자로 잘 알려진 MAP-kinase (erk1/2)의 인산화는 필요치 않음을 알 수 있었다. 흥미롭게도 액틴 중합 저해제인 cytochalasin D의 PMA와 공 처리는 오히려 세포부착을 PMA 단독 처리시 보다 증가시켰다. 또한 본 실험에서 사용된 SRB 염색법을 통한 세포부착 분석법은 최근 암 등 다양한 질환의 신약 표적 분자로 주목을 받고 있는 PKC 저해제의 초기 세포 기반 분석에 매우 유용하리라고 생각된다.

식물세포벽 가수분해효소 중 펙틴계효소에 대한 고찰 (Respection of Pectic Enzymes Among the Hydrolysis Enzymes of Plant Cell Wall)

  • 최동원;김인규
    • 한국식품영양학회지
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    • 제9권1호
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    • pp.92-98
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    • 1996
  • Pectic materials, which are widely spread in the plant cell wall as plant carbohydrates, plays a great role in food Industry that acts as a softening agent of fruits and vegetables, and gel forming agents. To study physiochemical properties and industrial applications of pectic enzymes that hydrolyzes pectin, classification, assay method and Industrial application are reviewed based on previous results.

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Virus-cell fusion inhibitory compounds from Ailanthus altissima Swingle

  • Lee, Hyang-Hee;Chang, Young-Su;Moon, Young-Hee;Woo, Eun-Rhan
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.264.1-264.1
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    • 2003
  • In order to search for the anti-HIV agents from natural products, Eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. (omitted)

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Antitumor Effects of Kluyveromyces marxianus TFM-7 Isolated from Kefir

  • Lee, Hyun-Jung;Nam, Bo-Ra;Kim, Jin-Man;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Food Science and Biotechnology
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    • 제16권1호
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    • pp.133-137
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    • 2007
  • The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

Genotoxicity Assessment of Erythritol by Using Short-term Assay

  • Chung, Young-Shin;Lee, Michael
    • Toxicological Research
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    • 제29권4호
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    • pp.249-255
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    • 2013
  • Erythritol is a sugar alcohol that is widely used as a natural sugar substitute. Thus, the safety of its usage is very important. In the present study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of erythritol. According to the OECD test guidelines, the maximum test dose was 5,000 ${\mu}g$/plate in bacterial reverse mutation tests, 5,000 ${\mu}g/ml$ in cell-based assays, and 5,000 mg/kg for in vivo testing. An Ames test did not reveal any positive results. No clastogenicity was observed in a chromosomal aberration test with CHL cells or an in vitro micronucleus test with L5178Y $tk^{+/-}$ cells. Erythritol induced a marginal increase of DNA damage at two high doses by 24 hr of exposure in a comet assay using L5178Y $tk^{+/-}$ cells. Additionally, in vivo micronucleus tests clearly demonstrated that oral administration of erythritol did not induce micronuclei formation of the bone marrow cells of male ICR mice. Taken together, our results indicate that erythritol is not mutagenic to bacterial cells and does not cause chromosomal damage in mammalian cells either in vitro or in vivo.

현지초(Geranium thunbergii) 추출물의 생리활성 및 Raw 264.7 cells에서의 항염활성 검증 (Verification of the Physiological Activity of Geranium thunbergii Extract and Anti-inflammatory Activity in Raw 264.7 Cells)

  • 박승미;오민정;이진영
    • 생명과학회지
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    • 제34권1호
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    • pp.28-36
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    • 2024
  • 본 연구에서는 현재까지 화장품 소재로써 다양한 연구가 진행되지 않은 현지초 추출물의 효능평가 및 항염 관련 활성 연구를 진행하였다. 현지초 추출물의 항산화능을 확인하기 위해 전자공여능 및 ABTS+ 라디칼 소거능을 측정한 결과, 각각 50 ㎍/ml의 농도에서 91%, 94%를 나타내어 항산화능이 우수함을 확인할 수 있었다. 미백활성 측정을 위해 tyrosinase 저해활성 측정을 실시하였으며 최고 농도인 1,000 ㎍/ml에서 24.8%의 저해능이 나타났다. 현지초 추출물의 주름개선 활성을 알아보기 위해 elastase 및 collagenase 저해활성 측정을 실시하였으며 그 결과, 각각 최고 농도인 1,000 ㎍/ml에서 30.6%, 90%의 저해능이 나타났고 collagenase 저해활성에서 우수한 저해능을 확인할 수 있었다. 세포 실험 진행을 위해 현지초 추출물 처리에 따른 대식세포 Raw 264.7의 생존율을 MTT assay에 의거하여 진행하였으며 100 ㎍/ml의 농도에서 83.6% 이상의 세포 생존율을 나타내어 이하의 세포 관련 실험 진행은 100 ㎍/ml 이하의 농도의 현지초 추출물을 가하여 실험을 실시하였다. NO assay에 의하여 현지초 추출물 처리에 따른 NO 생성 저해 활성을 측정한 결과, 100 ㎍/ml의 농도에서 74.9%의 저해율을 확인하였다. 단백질 발현 억제능을 알아보기 위해 western blot을 진행하였으며 현지초 추출물은 COX-2 및 iNOS 두 인자 모두 농도의존적으로 단백질 발현량이 저해됨을 확인할 수 있었다. 이러한 결과들에 의해 현지초 추출물은 항염 관련 기능성 화장품 소재로써 활용 가능성이 있다고 사료된다.

생물의약품 제조공정에서 마이코플라스마 정량 검출을 위한 TaqMan Probe Real-Time PCR (TaqMan Probe Real-Time PCR for Quantitative Detection of Mycoplasma during Manufacture of Biologics)

  • 이재일;김인섭
    • KSBB Journal
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    • 제29권5호
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    • pp.361-371
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    • 2014
  • Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants of biologic manufacturing processes. Conventional methods for mycoplasma testing, direct culture method and indirect indicator cell culture method, are lengthy, costly and less sensitive to noncultivable species. In this report, we describe a new TaqMan probe-based real-time PCR method for rapid and quantitative detection of mycoplasma contamination during manufacture of biologics. Universal mycoplasma primers were used for mycoplasma PCR and mycoplasma DNA was quantified by use of a specific TaqMan probe. Specificity, sensitivity, and robustness of the real-time PCR method was validated according to the European Pharmacopoeia. The validation results met required criteria to justify its use as a replacement for the culture method. The established real-time PCR assay was successfully applied to the detection of mycoplasma from human keratinocyte and mesenchymal stem cell as well as Vero cell lines artificially infected with mycoplasma. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of mycoplasma contamination during manufacture of biologics.

Statistical Optimization of the Lysis Agents for Gram-negative Bacterial Cells in a Microfluidic Device

  • Kim, Young-Bum;Park, Ji-Ho;Chang, Woo-Jin;Koo, Yoon-Mo;Kim, Eun-Ki;Kim, Jin-Hwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권4호
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    • pp.288-292
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    • 2006
  • Through statistically designed experiments, lysis agents were optimized to effectively disrupt bacterial cells in a microfluidic device. Most surfactants caused the efficient lysis of Gram-positive microbes, but not of Gram-negative bacteria. A Plackett-Burman design was used to select the components that increase the efficiency of the lysis of the Gram-negative bacteria Escherichia coli. Using this experimental design, both lysozyme and benzalkonium chloride were shown to significantly increase the cell lysis efficiency, and ATP was extracted in proportion to the lysis efficiency. Benzalkonium chloride affected the cell membrane physically, while lysozyme destroyed the cell wall, and the amount of ATP extracted increased through the synergistic interaction of these two components. The two-factor response-surface design method was used to determine the optimum concentrations of lysozyme and benzalkonium chloride, which were found to be 202 and 99 ppm, respectively. The lysis effect was further verified by microscopic observations in the microchannels. These results indicate that Gram-negative cells can be lysed efficiently in a microfluidic device, thereby allowing the rapid detection of bacterial cells using a bioluminescence-based assay of the released ATP.