The Unreliability of MTT Assay in the Cytotoxic Test of Primary Cultured Glioblastoma Cells
| Autor: | Yona Kim, Sun Ha Paek, Hwa Yeon Jo, Hyung Woo Park, Dong Gyu Kim, Hyo Eun Moon, Jin Wook Kim, Seongtae Bae |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
primary cell
Cell type medicine.diagnostic_test Tetrazolium salts Biology Molecular biology non-specific intracellular reduction Cellular and Molecular Neuroscience chemistry.chemical_compound chemistry Western blot Immunology medicine Cytotoxic T cell Original Article MTT assay ethanol Neurology (clinical) Primary cell Formazan Cytotoxicity Intracellular |
| Zdroj: | Experimental Neurobiology |
| ISSN: | 2093-8144 1226-2560 |
| DOI: | 10.5607/en.2015.24.3.235 |
| Popis: | MTT assay is commonly used to assess the cellular cytotoxicity caused by anticancer drugs in glioblastomas. However, there have been some reports insisting that MTT assay exhibited non-specific intracellular reduction of tetrazolium which led to underestimated results of cytotoxicity. Here, we examine whether or not MTT assay can lead to incorrect information regarding alcohol-induced cytotoxicity on immortalized and primary glioblastoma cells. MTT assay was applied to assess the ethanol-induced cytotoxicity at various ethanol concentrations. The cellular cytotoxicity induced by different doses of ethanol was analyzed and compared through several cytotoxic assays. Ethanol-induced cytotoxicity observed through MTT assay on both cell types was shown to be ethanol dose-dependent below a 3% concentration. However, the cytotoxicity was shown to be markedly underestimated only in primary cells at a 5% concentration. RT-PCR and Western Blot showed increased expressions of pro-apoptotic proteins and decreased expressions of anti-apoptotic proteins in an ethanol dose-dependent manner in both cell types. Furthermore, we present a possible mechanism for the unreliable result of MTT assay. A high concentration of ethanol induces more severe membrane damage and increased intracellular concentration of NADH in primary cells which enhances the nonspecific reduction of tetrazolium salt. Together, our findings demonstrate that the cytotoxicity on primary cells could inaccurately be assessed when detected through MTT assay. Therefore, a careful interpretation is needed when one would analyze the cytotoxic results of MTT assay, and it is suggested that other assays must be accompanied to produce more reliable and accurate cytotoxic results on primary glioblastoma cells. |
| Databáze: | OpenAIRE |
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