Simple and sensitive method for monitoring drug-induced cell injury in cultured cells

Journal Article · Sat Jun 01 04:00:00 EDT 1985 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(85)90290-8· OSTI ID:5148379

Shirhatti, V; Krishna, G

A simple, sensitive method has been developed for evaluating cell injury noninvasively in monolayer cells in culture. The cell ATP pool was radiolabeled by incubating the cells with (/sup 14/C)adenine. The uptake and incorporation of (/sup 14/C)adenine was shown to proportional to the number of cells. As determined by HPLC, about 65-70% of the incorporated /sup 14/C label was in the ATP pool, 15-20% was in the ADP pool, and the rest was in the 5'-AMP pool. When prelabeled cells were exposed to toxic drugs (acetaminophen, calcium ionophore A-23187, or daunomycin) there was a marked decrease in cell ATP with a concomitant increase in leakage of labeled nucleotides, mainly 5'-AMP and 5'IMP. The authors have shown that leakage of /sup 14/C label into the medium from the prelabeled cells may be employed for quantitation of cell injury. This new measure of toxicity was shown to correlate very well with LDH leakage from the cells, which is a well accepted measure of cell injury. The leakage of 5'-(/sup 14/C)AMP also correlated very well with the reduction of cell ATP in cardiac myocytes. This method has been used for monitoring drug-induced toxicity in liver cells, cardiac myocytes, and LB cells.

• Research Organization: National Heart, Lung, and Blood Institute, Bethesda, MD
• OSTI ID: 5148379
• Journal Information: Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 2; ISSN ANBCA
• Country of Publication: United States
• Language: English

Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ADENINES
ALKALINE EARTH METALS
AMINES
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIMETABOLITES
AROMATICS
ATP
AZAARENES
BODY
CALCIUM
CARBON 14 COMPOUNDS
CARDIOVASCULAR SYSTEM
CELL CULTURES
CHROMATOGRAPHY
DIGESTIVE SYSTEM
DRUGS
ELEMENTS
ENZYMES
GLANDS
HEART
HEMIACETAL DEHYDROGENASES
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
L CELLS
LABELLED COMPOUNDS
LACTATE DEHYDROGENASE
LIQUID COLUMN CHROMATOGRAPHY
LIVER
MAMMALS
METALS
MONITORING
MORPHOLOGICAL CHANGES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OXIDOREDUCTASES
PHENOL
PHENOLS
PREGNANCY
PURINES
RATS
RODENTS
SEPARATION PROCESSES
TOXICITY
TRACER TECHNIQUES
UPTAKE
VERTEBRATES