• 제목/요약/키워드: In vitro toxicology

검색결과 421건 처리시간 0.025초

Comparison of Sensitivity Between Balb/c 3T3 Cell and HaCaT Cell by NRU Assay to Predict Skin Phototoxicity Potential

  • Lee, Jong-Kwon;Lee, Eun-Hee;Lee, Sun-Hee
    • Toxicological Research
    • /
    • 제18권3호
    • /
    • pp.227-232
    • /
    • 2002
  • In order to find out the appropriate in vitro method for high correlation with in vivo, we com-pared the sensitivities of phototoxicity (PT) in vitro method between in human keratinocytes, HaCaT cells and in 3T3 fibroblast cells derived from Balb/c mice. Both cells were exposed to six known phototoxic chemicals : promethazine, neutral red, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen, or non-phototoxic chemical, ALS (ammonium laureth sulfate) and then irradiated with 5 J/$cm^2$ of UVA. Cell viability ($IC_{50}$ ) was measured by neutral red uptake (NRU) assay. The ratio of $IC_{50}$ value of chemicals in the presence and absence of UVA was determined by the cut-off value. The phototoxic potential of test chemicals in NRU assay was determined by measuring the photoirriation factor (PIF) with a cut-off value of 5. In both 3T3 and HaCaT cells, all known phototoxic chemicals were positive (over 5 of PIF value), except that bithionol was found to be non-phototoxic to HaCaT cells, and ALS, non-phototoxic chemical was negative. These results suggest that Balb/c 3T3 cell was more sensitive than HaCaT cell to predict phototoxicity potential.

The Study of Mutation Spectrum in Iac / Gene of Transgenic Big Blue$\textregistered$ Cell Line Following Short-Term Exposure to 4-Nitroquinoline N-oxide

  • Youn, Ji-Youn;Kim, Kyung-Ran;Cho, Kyung-Hea;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
    • /
    • 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
    • /
    • pp.64-64
    • /
    • 1996
  • Transgenic animal and cell line models which are recently developed in toxicology field combined with molecular biological technique, are powerful tools for studying of mutation in vivo and in vitro, respectively. The Big Blue mutagenesis assay system is one of the most widely used transgenic systems. Especially, for the study of direct acting mutagens, Big Blue cell line is very useful and powerful to evaluate mutagenicity because the mutation frequency and mutationspectrlun showed no distinct differences between cell line and animal. The Big Blue cell lines carry stably integrated copies of lambda shuttle vector containing lac I gene as a mutational target. These lambda shuttle vectors are useful for various mutagenesis related studies in eukaryotic system due to their ability to be exposed mutagen and then transfer a suitable target DNA sequence to it convenient organism for analysis. We tried to assess the mutagenic effect of 4-NQO with Big Blue cell line. After the treatment of 4-NQO, genomic DNA was isolated and lambda shuttle vector was packaged by in Vitro packaging and then these were plated on bacterial host in the presence of X-gal to screen mutation in the lac I. We determined MF as a ratio of blue plaques versus colorless plaques and now undergoing the mutation spectrum of 4-NQO in lac J gene sequence.

  • PDF

In vitro Alternatives to Skin Irritation Test

  • Shin, Dae-Sup;Kim, Dai-Byung;Ryu, Seung-Rel;Lee, Sun-Hee;Koh, Jae-Sook;Park, Won-Sae;Kim, Pu-Young
    • Biomolecules & Therapeutics
    • /
    • 제3권3호
    • /
    • pp.242-244
    • /
    • 1995
  • In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

  • PDF

Fullerene-C60의 포유류 배양세포를 이용한 염색체이상시험 (In vitro Mammalian Chromosomal Aberration Test of Fullerene-C60)

  • 김수진;임경택;조해원;한정희;김현영;양정선
    • Environmental Analysis Health and Toxicology
    • /
    • 제24권1호
    • /
    • pp.43-52
    • /
    • 2009
  • Fullerene의 유전독성을 평가하기 위하여 Chinese hamster유래의 난소유아세포(CHO-K1 cell)를 이용하여 직접법(-S9)과 대사활성화법(+S9 mix)의 염색체이상시험을 실시하였다. 시험물질은 1% CMC 나트륨염의 현탁액(1% CMC 용액)에 희석하여 조제하였다. 대사활성화를 시키지 않은 직접법의 염색체이상시험에서 24시간 투여군은 8단계의 농도(0.078, 0.156, 0.313, 0.625, 1.25, 2.5, 5, 10 mM)로 투여하여 실시하였다. 투여 농도 증가에 따른 염색체이상의 빈도가 증가하는 양상이 나타나지 않았다. 48시간의 투여군에서는 8단계의 농도(0.078, 0.156, 0.313, 0.625, 1.25, 2.5, 5, 10 mM)로 투여하여 실시하였는데 투여 농도 증가에 따른 염색체이상의 빈도가 증가하는 양상이 나타나지 않았다. 배수체의 염색체이상은 직접법에서 관찰되지 않았다. 대사활성화법을 이용하여 6시간 시험물질을 투여한 시험에 있어서는 8단계의 용량단계(0.078, 0.156, 0.313, 0.625, 1.25, 2.5, 5, 10mM)를 설정하였는데 투여 농도가 증가함에 따른 염색체이상빈도의 증가양상이 관찰되지 않았다. 이상의 결과를 종합할 때 본 시험물질은 본 시험 조건하에서 CHO-K1세포에서 대사활성화를 시켰을 때 염색체이상을 유발하지 않는 것으로 판단된다.

유전자수준에서 돌연변이 유발기전을 밝히는 Transgenic Mutagenesis Assay (Transgenic Mutagenesis Assay to Elucidaate the Mechanism of Mutation at Gene Level)

  • 류재천;윤지윤;조경혜;장일무
    • 한국환경성돌연변이발암원학회지
    • /
    • 제18권1호
    • /
    • pp.15-21
    • /
    • 1998
  • Transgenic animal and cell line models which are recently developed and used in toxicology fields combined with molecular biological technique, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Transgenic models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease processes. The lac I and lac Z gnee most widely used as a mutational target in transgenic systems. The assay is performed by treatment with putative mutagenic agents, isolation of genomic DNA from cells or tissues, exposure the isolated DNA to in vitro packaging extract, plating and sequencing. The results from these processes provide not only mutant frequency as quantitative evaluation but also mutational spectrum as qualitative evaluation of various agents. Therefore we introduce and review the principle, detailed procedure and application of transgenic mutagenesis assay system in toxicology fields especially in mutagenesis and carcinogenesis.

  • PDF

Toxic Mechanism of Atrazine in Transgenic Mutagenesis System using Big $Blue^{\textregistered}}$Rat2 lacI Transgenic Fibrohlasts And Hormonal Disturbances in vitro

  • Kim, Youn-Jung;Lee, Kilchul;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
    • /
    • 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
    • /
    • pp.128-128
    • /
    • 2001
  • Atrazine, one of herbicide widely used in agriculture, is classified as a possible human carcinogen (2B group) that may cause breast and ovarian cancers by IARC, and is known as an endocrine disrupter. Atrazine has been subjected to broad ranges of genotoxicity tests wi th predominantly negative results, but reported conflicting results across two or more independent tests as well. (omitted)

  • PDF

Estrogenic Activity Assessment of Alkylphenolic chemicals using in vitro assays : II. Competitive Receptor Binding Assay

  • Park, Hyo-Joung;Lee, Ho-Sa;Lee, Kilchul;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
    • /
    • 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
    • /
    • pp.126-126
    • /
    • 2001
  • Over the last few years, an increased awareness of endocrine disrupting chemicals (EDCs) and their potential to affect wildlife and humans has produced a demand for practical screening methods to identify endocrine activity in a wild range of environmental and industrial chemicals. It is clear that in vivo methods will be required to identify adverse effects produced by these chemicals. (omitted)

  • PDF

합성화학물질들의 유전독성평가(Ⅵ) -Chinese hamster lung세포를 이용한 17종 합성화학물질들의 염색체 이 상 시험 - (Evaluation of the Genetic Toxicity of Synthetic Chemicals (Ⅵ) -In vitro Chromosomal Aberration Assay with 17 Chemicals in Chinese Hamster Lung Cells -)

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Youn-Jung;Jeon, Hee-Kyung
    • Environmental Analysis Health and Toxicology
    • /
    • 제18권2호
    • /
    • pp.111-120
    • /
    • 2003
  • 합성화학물질들이 환경중에 유입되어 인체에는 물론 환경생태계에 많은 영향을 미치고 있어 이들의 유해성 검증은 매우 중요한 일이라 할 수 있다. 실제 산업체에서 사용되는 수많은 화학물질들의 유전적 손상 유발유무는 더욱이 중요한 일이라 할 수 있다. 이에 따라 산업체 공정과정에서 널리 사용되는 17종의 화학물질에 대해 Chinese hamster lung(CHL)세포를 이용한 염색체 이상 시험을 수행하여 이들이 염색체상에 구조적 이상을 유발하는지 관찰하였다. 그 결과, 본 연구에서 사용한 17종의 합성화학물질 중 2-nitroaniline(CAS No. 88-74-4)만이 대사활성화계 부재시 86.3 $\mu\textrm{g}$/ml의 농도에서 통계적으로 유의한 염색체 이상 유발능을 보였다. 반면 가장 높은 세포독성을 보인 1-chloroanthraquinone (CAS No. 82-44-0)은 0.8 ∼ 3.0 $\mu\textrm{g}$/ml의 시험농도범위에서 대사활성 존재 유무와 무관하게 염색체 이상을 유발하지 않았으며, 다른 15종의 물질들 역시 본시험 적용 농도 범위에서 염색체 이상 유발능을 관찰할 수 없었다.