• Title/Summary/Keyword: vero cell

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Effects of the Petroleum-ether Extract of Ginseng on the Cell Cycle and Protein Kinase C Activity in Cancer Cells (인삼 Petroleum-ether 추출물이 종양세포의 증식 주기 진행 및 Protein Kinase C의 활성에 미치는 영향)

  • 박민경;황우익
    • Journal of Ginseng Research
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    • v.20 no.3
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    • pp.219-225
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    • 1996
  • This study was performed to investigate the inhibition mechanism of cancer cell proof iferation caused by the petroleum-ether extract of ginseng against human rectum (HRT-18), colon (HT-29), llepatoma (Hep G2) and prostate (LNCaP) cancer cells and monkey kidney cells (Vero 76). Cells were treated with the petroleum-ether extract of ginseng (50 to 200 $\mu\textrm{g}$/ml) in G1 or S phase of the cell cycle, and proliferation and protein kinase C activity were measured. The petroleum-eth or extract of ginseng inhibited proliferation of HRT-18, HT-29, Hep G2 and LNCaP when treated in Gl phase, but not in S phase. This result shows that the ginseng extract arrests the cell cycle in G1 phase, resulting in the inhibition of cell proliferation. At the same concentrations, treatment of the ginseng extract in G1 phase decreased protein kinase C activity, while the treatment in S phase had no effect. This reault suggests that protein kinase C might be involved in the inhibition of the cell cycle and proliferation of cancer cells caused by the petroleum-ether extract of ginseng.

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Protective effect of methanol extract from citrus press cakes prepared by far-infrared radiation drying on $H_2O_2$-mediated oxidative damage in Vero cells

  • Wijesinghe, W.A.J.P.;Senevirathne, Mahinda;Oh, Myung-Cheol;Jeon, You-Jin
    • Nutrition Research and Practice
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    • v.5 no.5
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    • pp.389-395
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    • 2011
  • In the present study, a suitable drying method was developed for citrus press cakes (CPCs), which are produced as a by-product in citrus juice plants, and the protective effect of methanol extract of CPCs prepared by far-infrared radiation (FIR) drying against $H_2O_2$-induced DNA damage was evaluated versus that of freeze-dried CPCs. Methanol extract of FIR-dried CPCs exhibited comparatively good ROS scavenging activity versus the freeze-dried CPCs at the concentration of 100 ${\mu}g$/mL. The extract strongly enhanced the cell viability against $H_2O_2$-induced oxidative damage in Vero cells. Lipid peroxidation inhibitory activity of the extract from FIR-dried CPCs was comparable to that of the extract from freeze-dried CPCs. This sample also exhibited good protective effects against $H_2O_2$-mediated cell apoptosis as demonstrated by decreased apoptotic body formation in the nuclear staining with Hoechst 33342. In the comet assay, the CPC extracts exhibited strong inhibitory effects against $H_2O_2$-mediated DNA damage in a dose-dependent manner. Thus, this study demonstrated that FIR drying effectively preserves CPC as a functionally important natural antioxidant source and the FIR drying can be adapted for drying CPCs and is more economical for massive production than freeze drying.

Comparison of Embryonic Developmental Capacity by different Co-culture Time of Oocytes in IVF-ET Cycles (체외수정술에서 난자의 공배양 시점에 따른 배아 발생능력의 비교)

  • Lee, Hyun-Jung;Park, Kee-Sang;Song, Hai-Bum;Lee, Taek-Hoo;Cho, Young-Lae;Chun, Sang-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.21-28
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    • 2002
  • Objective: To evaluate whether co-culture of oocytes on vero cell monolayers from Day 0 (Day 0 group) after egg retrieval results in an increase in developmental capacity such as fertilization rate, embryo quality, blastulation and clinical pregnancy rate compared with co-culture of oocytes from Day 1 (Day 1 group). Methods: Sperms were treated with Hams F-10 supplemented with 10% human follicular fluid (hFF). Vero cells for co-culture were prepared in TCM-199 with 10% FBS. Oocytes were co-cultured from Day 0 and fertilized oocytes were co-cultured from Day 1 on vero cell monolayers in DMEM with 10% and 20% hFF, respectively after egg retrieval. On day 1, 2 and 5, fertilization rate and grade of embryos and blastocysts were evaluated. Results (fertilization rate, cleavage rate, grade of embryos and blastocysts and pregnancy rate) were considered statistically significant when p value was less than 0.05 using t-test and $x^2$. Results: In sibling oocytes of same cycles, no differences were found in fertilization rate (94.6 vs. 91.4%), cleavage rates (94.6 vs. 91.4%), embryo grade (on day 2 and 3) and blastulation (65.6 vs. 57.0%) and their grade. In different oocytes of different cycles (patients), no differences were found in fertilization (79.8 vs. 78.3%), cleavage rates (77.7 vs. 76.4%) and blastulation (56.0 vs. 45.3%), but pregnancy rate was higher in the Day 0 group than in the Day 1 group (60.0 vs. 42.9%). Conclusions: This study revealed that the embryonic development capacities were not affected by the different co-culture time in the sibling oocytes of same cycles. Although no statistical significance, because of small size of study, there was a trend for higher pregnancy rates in Day 0 group compared to Day 1 group in different oocytes of different cycles.

The Composition of the Root of Ixeris dentata var. albiflora Nakai. and Cell Viability and DPPH Radical Scavenging Activities of its Extract (흰씀바귀 (고채(苦菜), Ixeris dentata var. albiflora Nakai)뿌리의 성분 분석과 추출물의 세포 생존율 및 DPPH 라디칼 소거 활성)

  • Hong, Seul-Gi;Jeong, Dong-Myong;Kim, Ki-Young;Hwang, Eun-Hee
    • Journal of Nutrition and Health
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    • v.43 no.2
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    • pp.105-113
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    • 2010
  • Ixeris dentata var. albiflora Nakai, a herbal plant, is often used to make a strong stomach as an antiphlogistic used when dyspepsia and to improve appetite in Korea and China. And also it is used for adult diseases such as diabetes and liver diseases as Korean traditional medicine. In this study, the composition and DPPH radical scavenging activities of the root of Ixeris dentata var. albiflora Nakai and its effects on cell viability on vero and chang cells were investigated. Moisture, crude ash, crude protein and crude lipid were 79.14, 2.49, 8.28 and 2.56 g/100 g respectively. The highest mineral content was K. The major free sugars were glucose, fructose and sucrose. Major fatty acid are linoleic acid, palmic acid and linolenic acid. Major amino acids were glutamic acid, arginine and aspartic acid and the total contents of amino acids were 28.12 mg/g. The methanol extracts were further fractionated with n-hexane, chloroform, ethylacetate, butanol and water to get an active fraction. In addition, cell viabilities in each fraction were determined. Methanol extract, butanol, and aqueous fraction showed strong survival rates in vero cell and chang cell viability test, and hexane, chloroform, and ethylacetate fraction were examined for toxin in a cell. The root of Ixeris dentata var. albiflora Nakai had scavenging activities against DPPH radicals in a dose-dependent assay. Ethylacetate fraction's SC50 was $6.8\{\mu}g/mL$, very strong DPPH radical scavenging activities, but water fraction did not show any activity.

In vitro cytotoxic evaluation of some essential oils

  • P., Vijayan;Godavarthi, Ashok;Chandrashekhar, Raghu;Badami, Shrishilappa;SA, Dhanaraj;B., Suresh
    • Advances in Traditional Medicine
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    • v.3 no.4
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    • pp.187-190
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    • 2003
  • Seven essential oils were tested for in vitro cytotoxicity against the cancerous cell lines A-549, HEp-2 and DLA and normal BRL-3A, NRK-49F and Vero cell lines using standard MTT, SRB and dye exclusion techniques. The A-549 cell line was found to be the most susceptible to all the essential oils. The essential oils of A. nilagirica, A. calamus and O. sanctum were found to be the more active against these cells with mean $CTC_{50}$ values of 17.75, 19.00 and $24.37\;{\mu}g/ml$, respectively. The essential oil of Acorus calamus was found to be the most potent with low $CTC_{50}$ values against the cancerous and comparatively higher $CTC_{50}$ values against the normal cell lines. Artemisia pellens and Pelargonium graveolens oils also showed potent activity. These oils merit further investigation to identify the active principles and nature of the anti tumor activity in animal models.

Development of Serum Free Medium and Optimization of Porcine Rotavirus Vaccine Production

  • Ko, Yun-Mi;Kim, Myoung-Hwa;Kim, Min-Young;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.207-209
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    • 2005
  • Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final vaccine product. For porcine Rotavirus vaccine production, it is necessary to develop serum free medium which do not cause those problems. A new serum free medium was developed for porcine Rotavirus vaccine based on DMEM, and the performance of developed serum free medium was evaluated in terms of Vero cell growth and Rotavirus vaccine production. The cell density, gown in serum free medium developed, was similar with that in serum supplemented medium. Also, it was higher than that in other commercially available serum free medium. The productivity of Rotavirus vaccine using serum free medium developed and optimum production strategies will be also discussed.

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간암 치료 신약 개발 및 이의 제제화 연구(II. 제제화 연구)

  • 최명준;정홍석;장진수;황유경;안교한;이석종
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.65-65
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    • 1993
  • 봉합율의 최적 조건을 구하기 위해 다양한 방법으로 리포좀을 제조했으며, 인지질과 봉합 약물의 비는 20:1 이상이어야 하고, 리포좀 제조 방법에 따라 봉합율이 차이가 났으며, REV, REV를 freezing-thawing한 것과 SUV를 freezing-thawing한 리포좀에서 봉합율이 좋았다. REV를 freezing-thawing한 리포좀이 가장 좋았다 (66.7%, BPB; 115.7 ug trypsin/mg lipid). 콜레스테롤의 함량은 봉합율과 안정성을 고려하여 인지 질과의 비를 7.3으로 고정시켰다. 안정성은 투과성, light scattering, qucnching 등으로 측정했으며,4$^{\circ}C$,용액 상태에서 두 달 이상동안 보관했을 때 약 10% 정도가 리포좀 내에서 빠져나왔으며, 리포좀의 크기 변화는 거의 일어나지 않았다. Calcein봉합 리포좀을 Vero E6 cell에 투여했을 때, 매우 적은 양이 cell내로 들어가는 것을 형광 현미경으로 확인할 수 있었다.

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

  • Lee, Jae Il;Kim, In Seop
    • KSBB Journal
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    • v.29 no.5
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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.

Cytotoxic Effect of Aromatic and Aliphatic Compounds Produced by Streptomyces sp. Isolated in Korea (한국 Streptomyces SP.로부터 분리한 방향족 화합물과 지질 화합물의 세포독성 연구)

  • Shin, Suck-Woo;Ryeom, Kon
    • Biomolecules & Therapeutics
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    • v.5 no.2
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    • pp.215-221
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    • 1997
  • In an effort to screen new selective antitumor agents from the broth of soil microorganism, cytotoxicity oriented screening was performed against tumor cells and 3 compounds (Compound 1, 2 and 3) were isolated from Sreptomyces parvullus ISP 5048 and their chemical structures were determined. Among these compounds, Compound 2 showed the highest cytotoxicity against P388Dl and L1210. While the $IC_{50}$/ values of compound 2 against P388Dl and L1210 were 0.073$\mu$g/ml and 0.07$\mu$g/ml, respectively, and the $IC_{50}$/ value of Compound 3 was 0.17$\mu$g/ml against human lung cancer cells, A549, the cytotoxicity of Compound 2 and 3 against normal cell line, Vero E6 cell was about 4- and 8-fold lower than that of adriamycin. Based on the chemical analysis data, Compound 3 was octacosamicine A, a known antibiotic, which was reported by Dobasih et al. (1988). Taken together the results demonstrated that Compound 2 and Compound 3 has the possibility to be developed as antitumor agent because of its potent cytotoxicity as well as high selectivity against various cancer cell lines.

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Study of Cytotoxicity of an Actinomycete Isolated in Korea (토양에서 분리한 방선균의 세포 독성에 관한 연구)

  • Park, Joon-Koo;Choi, Boung-Don;Kim, Seung-Chul;Ryeom, Kon
    • Environmental Analysis Health and Toxicology
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    • v.8 no.3_4
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    • pp.7-12
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    • 1993
  • An Actinomycete strain isolated from Mt. Dea-Dun had a strong antifungal activity. The culture brith produced by isolated strain showed only antifungal activity against fungi with the exception of yeast and bacteria. It was heat stable, dissolved in ehtylacetate. The concentrated antifungal agent showed cytotoxicity against HEP-2 and HeLa as tumor cell line, and showed weak cytotoxicity against VERO 36 as normal cell line. Morphological and physiological characteristics were tested with isolated strain. The spore color of isolated strain was gray. It had a short chain and produced brown colored lytic substance in yeast extract-malt agar. The cell wall of isolated strain was composed of meso-DAP, and we suggested it as genus Actinomadura. In the existing of chemical inhibitor, isolated strain grew on the condition of 0.0001% crystal violet, 0.1% phenol, 0.01% sodium azide and 10% sodium chloride. Carbon utilization of isolated strain was shown that glucose, sucrose, manitol and sodium citrate were well utilized.

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