• Title/Summary/Keyword: XTT

Search Result 140, Processing Time 0.028 seconds

전남지역 생활하수처리장과 농공단지폐수처리장 방류수에 대한 세포독성 평가 - XTT, SRB in vitro bio-assay를 이용한 -

  • Lee, Moon-Hee;Han, Sang-Guk
    • Proceedings of the Korean Environmental Sciences Society Conference
    • /
    • 2005.11a
    • /
    • pp.185-188
    • /
    • 2005
  • XTT assay와 SRB assay를 이용하여 Hep3B와 L929 세포에서 생활하수처리장과 농공단지폐수처리장 방류수의 세포독성을 알아본 결과 다음과 같은 결론을 얻었다. 1) XTT, SRB 실험법에 적용하여 대표적인 발암성 물질인 Benzo(a)Pyrene의 세포독성을 평가 할 수 있었다. Benzo(a)Pyrene 30uM농도는 Hep 3B와 L929 세포에서 약 50%의 세포독성을 나타내었다. 2) 단백질합성과 세포호흡의 활성도로 세포독성을 평가할 수 있는 실험법인 XTT와 SRB 실험법에 적용하여 방류수를 추출 농축한 실제 시료에 대한 세포독성을 평가할 수 있었다. 3) 생활하수처리장의 방류수에 비해 다종 고농도의 화학물질로 오염되어있는 농공단지폐수처리장 방류수에서의 독성발현이 더 높았다. 4) 생활하수처리장 방류수와 농공단지폐수처리장 방류수 중 오염도가 가장 심각한 4지점의 화학물질 분석 결과와 in vitro bio-assay에 의한 세포독성 결과가 일치하였다.

  • PDF

Toxic Effect on Phenolic Compound by Colorimeteric Assay in Normal NIH 3T3 Fibroblasts

  • Jin Byung-Jo;Lee Joo-Hyun;Choi Ki-Wook;Lee Jae-Kyoo;Han Du-Seok
    • Biomedical Science Letters
    • /
    • v.10 no.3
    • /
    • pp.263-268
    • /
    • 2004
  • This study was carried out to evaluate the cytotoxic effect of phenolic compound on normal NIH 3T3 fibrolasts. The colorimetric assay for phenol compound, syringic acid was performed by MTT assay or XTT assay. MTT or XTT assays are known as a very sensitive method in measuring the cytotoxic effect of chemical agents in vitro. In the present study, syringic acid on normal Nlli 3T3 fibroblasts did not show any cytotoxicity for MTT assay or XTT assay compared with control after cells were treated with various concentrations of syringic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 3,340.9 μM and 2,462.4 μM of syringic acid, respectively. From the above the results, it is suggested that phenolic compound of syringic acid did not have any cytotoxicity on normal NIH 3T3 fibroblasts.

  • PDF

The Inhibitory Effect of Ferulic Acid and Related Phenolic Compounds against Cancer Cell Lines (Ferulic Acid와 관련 페놀화합물의 암세포주에 대한 독성억제효과)

  • Han, Du Seok;Chun, Joo Won;Jeon, Sung Woo;Ba다, Seung Hwa
    • YAKHAK HOEJI
    • /
    • v.49 no.5
    • /
    • pp.365-369
    • /
    • 2005
  • The inhibitory effect of ferulic acid and related phenolic compounds on the growth of normal cell lines and can­cer cell line was evaluated by the MTT and XTT methods. Ferulic acid decreased the cell viability of human skin melanoma cells by the MTT method and the cell adhesion activity of human oral epithelioid carcinoma cells by the XTT method. These results suggest that ferulic acid has a potential anticancer activity.

In Vitro Bioassay for Transforming Growth Factor-$\beta$ Using XTT Method

  • Kim, Mi-Sung;Ahn, Seong-Min;Moon, Aree
    • Archives of Pharmacal Research
    • /
    • v.25 no.6
    • /
    • pp.903-909
    • /
    • 2002
  • Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

Effect of Ferulic Acid on Cell Viability and Cell Adhesion Activity in Normal Human Gingival Fibroblasts

  • Lee Joo-Hyun;Jin Byung-Jo;Son Il-Hong;Han Du-Seok
    • Biomedical Science Letters
    • /
    • v.10 no.3
    • /
    • pp.269-273
    • /
    • 2004
  • This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferolic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferolic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 2,130.0 μM and 1,773.7 μM ferolic acid, respectively. These results suggest that ferolic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

  • PDF

Effect of Kaempferol on the Cytotoxicity Induced Oxygen Free Radicals in Skin Fibroblast Derived from Human In Vitro

  • Lee, Jai-Kyoo;Ha, Dae-Ho
    • Biomedical Science Letters
    • /
    • v.14 no.3
    • /
    • pp.193-198
    • /
    • 2008
  • In order to evaluate on the effect of kaempferol on the cytotoxicity of oxygen tree radicals, XTT assay was performed to determine the cell viability after skin fibroblasts derived from human (Detroit 51) that were treated with various concentrations of hydrogen peroxide $(H_2O_2)$. And also, the effect of kaempferol on the cytotoxicity induced by H202 that was examined by cell viability, lactate dehydrogenase (LDH) activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. $H_2O_2$ decreased cell viability in dose-dependent manner in these cultures and the $XTT_{90}\;and\;XTT_{50}$ values were determined at concentration of $35{\mu}M\;and\;90{\mu}M$ of $H_2O_2$ after skin fibroblasts derived from human were treated with $15{\sim}90{\mu}M$ of $H_2O_2$ for 6 hours, respectively. $H_2O_2$ was highly toxic on cultured skin fibroblasts derived from human by toxic criteria of Brenfreund and Puerner (1984). In the protective effect of kaempferol on $H_2O_2$-induced cytotoxicity, kaempferol increased DPPH radical scavenging activity and significantly decreased LDH activity. From these results, it is suggested that oxygen tree radical, $H_2O_2$, was highly toxic on cultured skin fibroblasts derived from human, and also kaempferol of flavonoid showed the protection on $H_2O_2$-induced cytotoxicity.

  • PDF

Effect of Cadmium on $C_6$ Glioma Cells in Culture

  • Son Young-Woo;Lee Kang-Chang
    • Biomedical Science Letters
    • /
    • v.12 no.3
    • /
    • pp.275-279
    • /
    • 2006
  • It is demonstrated that cadmium has cytotoxic effect on glial cells, oxygen radicals are involved in cadmium-induced cytotoxicity. However, the toxic mechanism of cadmium is left unknown so far. The purpose of this study was to examine the cytotoxicity of $CdCl_2$ on $C_6$ glioma cells. The cytotoxicy was measured by cell viability via XTT assay in $C_6$ glioma cells. Colorimetric assay such as XTT assay is regarded as a very sensitive screening method for the determination of the cell viability on a lots of chemicals. In this study, $CdCl_2$ decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were treated with various concentrations of $CdCl_2$ for 48 hours. $IC_{90}\;and\;IC_{50}$ values for XTT assay was determined at $5{\mu}M\;and\;55{\mu}M$ of $CdCl_2$, respectively. These results suggest that $CdCl_2$ has highly cytotoxic effect on $C_6$glioma cells by the decrease of cell viability.

  • PDF

Cytotoxic Effect of Syringic Acid on Human Oral Epithelioid Carcinoma Cells

  • Lee Joo-Hyun;Han Du-Suk;Jekal Seung-Joo;Lee Jae-Hyung;Kim Chong-Ho;Yoo Min;Park Seung-Taeck
    • Biomedical Science Letters
    • /
    • v.11 no.3
    • /
    • pp.337-341
    • /
    • 2005
  • This study was undertaken to clerify the cytotoxic effect of syringic acid by colorimetric assay on human cancer cells. For the evaluation of cytotoxicity of syringic acid, the cell viability and cell adhesion activity of syringic acid on cancer cells, human oral epithelioid carcinoma cells were determined using by colorimetric assays such as MTT (3-[4,5­dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and XTT (2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]­2H-tetrazolium-5-caboxanilide) assay, respectively after human oral epithelioid carcinoma cells were treated with syringic acid for 48 hours. In this study, the cell viability of syringic acid on human oral epithelioid carcinoma cells showed a significant decrease by MTT assay compared with control, and also, the cell adhesion activity by XTT assay was decreased significantly in these cells after cells were treated with various concentrations of syringic acid for 48 hours. $MTT_{50}\;and\;XTT_{50}\;were\;282.3\;{\mu}M\;and\;418.8{\mu}M$ syringic acid, respectively. These results suggest that syringic acid shows midcytotoxic effect on human oral epithelioid carcinoma cells by the decreasement of the cell viability and the cell adehision activity assessed by colorimetric assay in these cultures.

  • PDF

Effect of Persimmon Leaves Extract on the Melanogenesis and Cell Viability in Cultured Melanoma Cells Injured by Reactive Oxygen Species (시엽추출물이 활성산소로 손상된 멜라닌세포종의 멜라닌합성 및 세포생존율에 미치는 영향)

  • Ha, Dae-Ho;Lee, Jae-Kyoo;Choi, Yu-Sun
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.22 no.5
    • /
    • pp.1304-1308
    • /
    • 2008
  • This study was performed to evaluate the effect of persimmon leaves extract on the reactive oxygen species (ROS) in cultured melanoma cells. The B16/F10 melanoma cells were treated with various concentrations of t-butyl hydroperoxide (t-BHP). And also, the effect of persimmon leaves (PL) extract on the cytotoxicity mediated by t-BHP was done on the cell viability, tyrosinase activity and melanogenesis by colorimetric assays. In this study, t-BHP decreased cell viability in dose-dependent manner and XTT90 and XTT50 values were measured at 10 and 35 uM of PL, respectively in these culture. And also, XTT50 value was assessed as a highly toxic effect on cultured melanoma cells by the toxic criteria. In the effect of PL extract on the t-BHP-mediated cytotoxicity, PL extract significantly increased the cell viability injured by t-BHP in cultured B16/F10 melanoma cells. PL also showed the decreased tyrosinase activity and melanogenesis. From these results, it is suggested that ROS such as t-BHP showed highly toxic effect on cultured melanoma cells, and also, PL extract inhibited the tyrosinase activity and melanogenesis in cultured melanoma cells injured by ROS.

The Cytotoxic Activity of 3,4,5-Trihydroxybenzoic Acid Methylester and Related Compounds against Skin and Oral Cancer Cell Lines (3,4,5-Trihydroxybenzoic Acid Methylester와 관련 화합물의 피부암 및 구강암 세포주에 대한 세포독성)

  • Lee, Jae-Sug;Han, Du-Seok;Kang, Jeong-Il;Baek, Jong-Min;Baek, Seung-Hwa
    • YAKHAK HOEJI
    • /
    • v.54 no.2
    • /
    • pp.112-121
    • /
    • 2010
  • The cytotoxic activity of 33,4,5-trihydroxybenzoic acid methylester and related compounds on the growth of normal cell lines, human skin melanoma cells and human oral epithelioid cell line were evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis-[2-methoxy-4-nitro-5-sulfo-phenyl]-2-H-tetrazolium-5-caboxanilide (XTT) methods. 3,4,5-Trihydroxybenzoic acid methylester decreased the cell viability of human skin melanoma cells and human oral epithelioid cells shown by the MTT method and the cell adhesion activity of human skin melanoma cells and human oral epithelioid cells shown by the XTT method. In light microscopy, 100 ${\mu}M$ 3,4,5-trihydroxybenzoic acid methylester showed the highest cytotoxic activity. These results suggest that 3,4,5-trihydroxybenzoic acid methylester has a potential anticancer activity.