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Differential Sensitivities of Cytotoxicity Assays on C6 Glioma Cells Exposed to Dinitrobenzenes
Author(s) -
Rivera Berenise,
Miller Richard Timothy
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.838.1
Subject(s) - cytotoxicity , glioma , viability assay , neutral red , chemistry , cytotoxic t cell , mtt assay , in vitro , microbiology and biotechnology , biophysics , biochemistry , biology , cancer research
Dinitrobenzene (DNB) refers to a mixture of three isomers: 1,2‐, 1,3‐ and 1,4‐DNB. These DNBs are used as common intermediates in the dye and plastic industries. 1,2‐ and 1,4‐DNB cause methemoglobinemia, in contrast 1,3‐DNB is toxic to cerebellum and certain brainstem nuclei in rats. To our knowledge, no studies have been conducted to determine if the cytotoxic effects of 1,2‐ and 1,4‐DNB parallel those of 1,3‐DNB in neuronal or glial cell models. The purpose of this in vitro study was to compare the relative ability of different cytotoxicity assays to detect early changes in cell viability. C6 Glioma cells were exposed to 10–25 μM DNBs for up to 6 hours. Cytotoxicity was then determined using MTT reduction for mitochondrial integrity, and neutral red (NR) uptake for lysosomal integrity. C6 Glioma cells exhibited a concentration‐ and time‐dependent decrease in NR uptake following a four hour exposure to 1,2‐ and 1,4‐DNB. Effects were most evident between 50–250 μM (80% and 8%, respectively). Under the same conditions, no evidence for NR uptake was observed upon exposure of C6 glioma cells to 1,3‐DNB. In contrast to results obtained with NR, no reduction of MTT by C6 glioma cells was observed at any time or concentration used in these studies. These preliminary data indicate that under the conditions used in our system, NR is a more sensitive indicator of changes in cell viability. This project was supported in‐part by 5G12‐RR008124 from NCRR at UTEP and by NIEHS/NIH grant ES011982 to RTM.