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(프로토콜) Skin Corrosion (ECVAM Validated)
 

 
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Skin Corrosion (ECVAM Validated)

 

The Model

MatTek's patented EpiDerm™ System consists of normal, human-derived epidermal keratinocytes (NHEK) which have been cultured to form a multilayered, highly differentiated model of the human epidermis. These "ready-to-use" tissues, also known generically as Normal Human-3D (NHu-3D), are cultured on specially prepared cell culture inserts using serum free medium, attain levels of differentiation on the cutting edge of in vitro skin technology. Ultrastructurally, the EpiDerm Skin Model closely parallels human skin, thus providing a useful in vitro means to assess dermal irritancy and toxicology. For more information on the EpiDerm model, click here.

The Method

프로토콜 요청은 이곳으로.

  1. Transfer tissues from agarose to assay medium

  2. Incubate (37 ±1°C, 5 ±1% CO2, 95% RH) overnight (18 ±3 h)

  3. Dose tissues with 30 μl/25 mg TS, PC, or NC

  4. Expose one set of tissues for 3 min (37 ±1°C, 5 ±1% CO2, 95% RH) and a second set of tissue for 60 min (37 ±1°C, 5 ±1% CO2, 95% RH)

  5. Stop exposure by rinsing in DPBS

  6. Blot tissues and perform MTT assay

  7. Read OD in a plate spectrophotometer at 550-570nm

 

The Endpoints

MTT Tissue Viability Assay

 

 

Regulatory Acceptance

European Centre for the Validation of Alternative Methods (ECVAM)

OECD TG 431

 

Technical References

 

693. CLASSIFICATION AND LABELING OF INDUSTRIAL PRODUCTS WITH EXTREME PH BY MAKING USE OF IN VITRO METHODS FOR THE ASSESSMENT OF SKIN AND EYE IRRITATION AND CORROSION IN A WEIGHT OF EVIDENCE APPROACH. Scheel1, J., Heppenheimer2,A., Lehringer1, E., Kreutz1, J., Poth2, A., Ammann1, H., Reisinger1, K., Banduhn1, N.  1Henkel AG & Co. KGaA, Düsseldorf, Germany. 2Harlan Cytotest Cell Research GmbH, Rossdorf, Germany. Toxicology in Vitro, 25, 1435–1447 (2011).

674. RETROSPECTIVE ANALYSIS OF THE EPIDERM 3-MINUTE PREDICTION MODEL FOR ASSESSMENT OF GHS SKIN CORROSION PACKING GROUP SUB-CATEGORY 1A. Kandarova, H., Kaluzhny, Y., Kubilus, J. Hayden, P., Sheasgreen, J. and Klausner, M.MatTek Corporation, Ashland, MA, USA MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia. Presented at 8th World Congress on Alternatives and Animal Use, Montreal, Canada, 2011. 

582. NON-ANIMAL METHODS FOR TOXICITY TESTING: SKIN IRRITATION/CORROSION. AltTox.org, 2008

565. FURTHER DEVELOPMENT OF AN EPIDERM™ IN VITRO SKIN IRRITATION TEST FOR THE GLOBALLY HARMONIZED SYSTEM (GHS-SIT) OF CLASSIFICATION AND LABELING OF CHEMICALS. Armento, A., Kandarova, H., Klausner, M. and Hayden, P.J. MatTek Corporation, Ashland, Massachusetts, USA. Presented at 49th Annual SOT Meeting, Salt Lake City, Utah (2010).

491. DEVELOPMENT OF AN IN VITRO CORROSION/IRRITATION PREDICTION ASSAY USING THE EPIDERM™ SKIN MODEL. Kidd1,2, D.A., Johnson1, M., Clements1, J. 1Department of Genetic and Molecular Toxicology, Covance Laboratories Ltd., Otley Road, Harrogate, North Yorkshire HG3 1PY, UK. 2Department of Biology, University of York, Heslington, York, North Yorkshire YO1 7EP, UK. Toxicology In Vitro, 21, 1292-1297 (2007).

 

 

 

 

 

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