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Titolo:
Evaluation of hepatic subcellular fractions for Alamar blue and MTT reductase activity
Autore:
Gonzalez, RJ; Tarloff, JB;
Indirizzi:
Univ Sci Philadelphia, Dept Pharmaceut Sci, Philadelphia, PA 19104 USA Univ Sci Philadelphia Philadelphia PA USA 19104 hiladelphia, PA 19104 USA
Titolo Testata:
TOXICOLOGY IN VITRO
fascicolo: 3, volume: 15, anno: 2001,
pagine: 257 - 259
SICI:
0887-2333(200106)15:3<257:EOHSFF>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
COLORIMETRIC ASSAY; PROLIFERATION; TOXICITY; DYE;
Keywords:
Alamar blue; MTT; enzymatic reduction; cell viability;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
10
Recensione:
Indirizzi per estratti:
Indirizzo: Tarloff, JB Univ Sci Philadelphia, Dept Pharmaceut Sci, 600 S 43rd St, Philadelphia, PA 19104 USA Univ Sci Philadelphia 600 S 43rd St Philadelphia PAUSA 19104
Citazione:
R.J. Gonzalez e J.B. Tarloff, "Evaluation of hepatic subcellular fractions for Alamar blue and MTT reductase activity", TOX VITRO, 15(3), 2001, pp. 257-259

Abstract

Alamar blue and MTT are indicators used to measure cytotoxicity of variouschemicals in cultured cells. Both Alamar blue and MTT are reduced by mitochondrial enzymes. We observed enhanced fluorescence of Alamar blue when kidney epithelial cells were co-incubated with hepatic post-mitochondrial supernatant (S9) fractions as compared with cells incubated in the absence of S9 fractions. The present studies were carried out to determine whether hepatic cytosolic and/or microsomal enzymes were capable of metabolizing Alamarblue and/or MTT to their reduced products. Livers from female Sprague-Dawley rats were used to prepare S9 fraction, and mitochondrial, microsomal andcytosolic fractions. Fractions containing 1 or 5 mg protein/ml were incubated with Alamar blue or MTT for up to 4 h. Fluorescence (Alamar blue) or absorbance (MTT) were determined and expressed as differences between treatedwells and controls. Hepatic fractions (S9, mitochondria, microsomes and cytosol) caused concentration- and time-dependent increases in Alamar blue fluorescence and MTT absorbance. Reduction of Alamar blue and MTT by hepatic S9 fraction was abolished by heating. Reduction of Alamar blue by hepatic mitochondria was approximately equivalent to that catalyzed by hepatic S9 fraction or cytosol. Reduction of MTT by hepatic mitochondria was approximately equivalent to that catalyzed by hepatic S9 fraction or microsomes. Thesedata indicate that mitochondrial, cytosolic and microsomal enzymes reduce Alamar blue and MTT. Therefore, caution should be exercised in ascribing decreases in viability as due solely to mitochondrial damage when using either of these dyes. (C) 2001 Elsevier Science Ltd. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 12/07/20 alle ore 12:23:37