[The utility of MTT and XTT colorimetric tests in the studies conducted in vitro with Toxoplasma gondii tachyzoites].

Wiadomosci parazytologiczne Pub Date : 2010-01-01
Katarzyna Dzitko, Dominika Dudzińska, Marcin Grzybowski, Henryka Długońska
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Abstract

Tetrazolium salts are widely used as indicators of metabolic activity for both eukaryotic and prokaryotic cells. Live cells reduce the tetrazole ring in MTT or XTT salts and then a colored formazane formed can be assessed spectrophotometrically. Despite widespread use of MTT/XTT reduction tests biochemical mechanisms of the reaction are still unknown, and each test application case requires standardization of experimental conditions. In the present study we tested in vitro the utility of both MTT and XTT salts to determine the influence of selected extracellular agents for T. gondii tachyzoites and their host cells (i.e. mouse L929 fibroblasts). The results showed that MTT is reduced more intensively than XTT by host and parasite cells. The attenuation of T. gondii tachyzoites resulted in a decrease of reduction level of both tetrazolium salts, particularly of XTT. Using MTT we found also that T. gondii is not susceptible to extremely toxic substance, sodium azide. Our results confirmed a high usefulness of MTT reduction tests in numerous studies on eukaryotic cells.

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[MTT和XTT比色法在刚地弓形虫速殖子体外研究中的应用]
四氮唑盐被广泛用作真核和原核细胞代谢活性的指示器。活细胞在MTT或XTT盐中还原四唑环,然后形成彩色的甲酸,可以用分光光度法测定。尽管广泛使用MTT/XTT还原试验,但其反应的生化机制尚不清楚,并且每个试验应用案例都需要标准化实验条件。在本研究中,我们在体外测试了MTT和XTT盐的效用,以确定选定的细胞外药物对弓形虫速殖子及其宿主细胞(即小鼠L929成纤维细胞)的影响。结果表明,宿主和寄生虫细胞对MTT的还原比XTT更强烈。弓形虫速殖子的衰减导致两种四氮唑盐的还原水平下降,尤其是XTT。利用MTT,我们还发现弓形虫对极毒物质叠氮化钠不敏感。我们的结果证实了MTT还原试验在真核细胞的大量研究中具有很高的实用性。
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