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

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 proph...

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Bibliographic Details
Main Authors: Guzman, Juantia J, Gross, Clark L, Smith, William J, Kelly, Susan A
Format: Report
Language:English
Subjects:
ASSAYING
Biochemistry
BLOOD
CELLS(BIOLOGY)
Chemical, Biological and Radiological Warfare
CYTOTOXICITY ASSAYS
CYTOTOXINS
DRUGS
EPIDERMIS
EXPOSURE(PHYSIOLOGY)
FLUORESCENT DYES
HD AGENT
HEK(HUMAN EPIDERMAL KERATINOCYTES)
LYMPHOCYTES
OXIDATION REDUCTION REACTIONS
PE62384A
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Summary: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.