Determination of viability of primary glioblastoma cell culture under the influence of various concentrations of endothelial monocyte-activating polypeptide II (EMAP II) using trypan blue staining

DOI: https://doi.org/10.7124/FEEO.v37.1772
Keywords: cytokine EMAP II, primary glioma cell culture, cytotoxicity, trypan blue dye exclusion test

Abstract

Aim. Confirmation of the previously established (by MTT assay) cytotoxic potential of the EMAP II polypeptide against primary glioblastoma cell culture using an independent method of trypan blue exclusion test, which is widely used for selective cell staining. Methods. Primary cell culture obtained from malignant glioma tissue fragments after surgery was treated with EMAP II at various concentrations. The trypan blue dye exclusion test was used to determine the number of viable cells present in the cell suspension. In this case, living cells, which have intact cell membranes, remain transparent due to the exclusion of the dye, while dead cells absorb the dye and turn blue. Results. A dose-dependent cytotoxic effect of EMAP II was recorded, which was manifested in the range of high drug concentrations 10 μM, 2 μM, 640 pM. Conclusions. The trypan blue exclusion test, due to its simplicity and availability, can be used for preliminary screening studies of the effect of EMAP II on malignant glioma cells, but it is significantly inferior to the MTT-test in sensitivity and informativeness.

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