In Vitro Cytotoxicity Assays of Human Epidermal Keratinocytes in Culture Exposed to Sulfur Mustard

Abstract

A number of studies have described several in vitro cytotoxicity assays that are convenient methods for assessing cell viability. Viability assessment of human peripheral blood lymphocytes (PBL) by flow cytometry of propidium iodide (PI) exclusion has been a useful tool for screening candidate prophylactic (PI) as well as therapeutic (T) drugs for medical countermeasures against sulfur mustard (HD)) for many years at our institute. Our laboratory has reported problems associated with simple vital dye uptake as a measure of cytotoxicity in human epidermal keratinocytes (HEK) following in vitro exposure to HD. We have evaluated five alternative assays of cell viability for HEK exposed to HI) in vitro: Calcein-AM, a fluorescent dye; Alamar Blue, a redox fluorescent dye; Neutral Red, a chromogenic dye; Methyl tetrazolium salt (MTS), a colorimetric assay; and viable cell number. At the highest HI) dose tested, the viable cell number, Calcein-AM, Alamar Blue and Neutral Red assays were more sensitive than propidium iodide uptake.

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Document Details

Document Type
Technical Report
Publication Date
Jan 01, 2000
Accession Number
ADA385183

Entities

People

  • Clark L. Gross
  • Juantia J. Guzman
  • Susan A. Kelly
  • William J. Smith

Organizations

  • United States Army Medical Research Institute of Chemical Defense

Tags

DTIC Thesaurus Topics

  • Acids
  • Biological Sciences
  • Biomedical Research
  • Blood
  • Cells
  • Chemical Warfare
  • Chemical Warfare Agents
  • Chemical Weapons
  • Culture Techniques
  • Dyes
  • Epithelial Cells
  • Fluorescent Dyes
  • Indicator Dyes
  • Viability

Fields of Study

  • Biology
  • Medicine

Readers

  • Analytical Chemistry
  • Geochemistry
  • Molecular Genetics