Different viabilities and toxicity types after 6-OHDA and Ara-C exposure evaluated by four assays in five cell lines

• Published in: Toxicology in vitro Vol. 22; no. 1; pp. 182 - 189
• Main Authors: Puttonen, Katja A., Lehtonen, Šárka, Lampela, Pasi, Männistö, Pekka T., Raasmaja, Atso
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-02-2008
• Subjects: Animals; Antimetabolites, Antineoplastic - administration & dosage; Antimetabolites, Antineoplastic - toxicity; Apoptosis; Apoptosis - drug effects; Cell death; Cell Death - drug effects; Cell Line; Cell Survival - drug effects; CHO Cells; Cricetinae; Cricetulus; Cytarabine - administration & dosage; Cytarabine - toxicity; Dose-Response Relationship, Drug; Haplorhini; Humans; Necrosis; Oxidopamine - administration & dosage; Oxidopamine - toxicity; Rabbits; Rats; Toxicity Tests - methods; Viability assay; SH-SY5Y; Viability assay; SMC; 6-OHDA; CHO; MTT; Ara-C; Necrosis; C6; Cell death; CV1-P; PI; Apoptosis
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2007.07.005

Cell viability studies are useful when screening novel drugs for the diseases that are related to either increased cell death or enhanced cell survival. There are numerous assays but the results that they produce are rarely unanimous. Here we compared the performance of (1) morphological microscopic assay with double DNA staining, (2) propidium iodide–digitonin assay, (3) MTT-assay, and (4) ATP-assay in human neuroblastoma (SH-SY5Y), rat glioma (C6), rabbit smooth muscle (SMC), Chinese hamster ovary (CHO) and monkey fibroblast cells (CV1-P) exposed to cytosine arabinoside (Ara-C) and 6-hydroxydopamine (6-OHDA). We found that neuronal SH-SY5Y cells were most sensitive to both toxins and the results in all viability tests correlated well. All the other cell lines were much more resistant, particularly to Ara-C but also to 6-OHDA. Toxicity of the compounds was best revealed by MTT and ATP assays, measuring the metabolic activity of the cells, and only occasionally by morphological observations or with the propidium iodide–digitonin assay which is based on the cell membrane integrity. In this research, Ara-C induced pure apoptosis whereas the toxicity type of 6-OHDA was dose-dependent. The use of several viability tests and cell lines is recommended when studying cell death, particularly apoptosis, and performance of antiapoptotic compounds.