Mitochondrial and nonmitochondrial reduction of MTT: Interaction of MTT with TMRE, JC‐1, and NAO mitochondrial fluorescent probes

Bernaś, Tytus; Dobrucki, Jurek

doi:10.1002/cyto.10080

Cited by 263 publications

(163 citation statements)

References 32 publications

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“…Bernas and Dobrucki (2002) have also shown that MTT reduction may be catalyzed by a number of nonmitochondrial enzymes. In their study, only 25-45% of MTT-formazan was associated with mitochondria after 25 min of incubation and most MTT-formazan deposits were not coincident with mitochondria.…”

Section: Discussionmentioning

confidence: 99%

Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells

et al. 2006

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The MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay is widely accepted as a simple and reproducible method for determining cell proliferation or cytotoxicity in vitro. In this study, we show that the flavonoids quercetin, rutin and luteolin but not apigenin can reduce MTT in the absence of live cells in the following order: quercetin >> rutin > luteolin > apigenin. Moreover, this reduction can be influenced by medium type and serum. The final concentrations of the flavonoids used were 200, 100, 50, 25 and 12.5 lg/mL. MTT reduction in Dulbecco's Modified Eagle's Medium (DMEM) is statistically higher than those in RPMI 1640 and F12 media, which are generally similar. Particularly for luteolin, MTT reduction is considerably higher with serum than without serum. In the case of quercetin at 50 lg/mL, a serum concentration of even only 0.01% is sufficient to significantly enhance MTT reduction versus that at 0% (P < 0.05). Serum at concentrations ranging from 0% to 5% also dosedependently affects the pattern of formazan crystal formation. In the presence of 0.156-5% serum, the formazan crystals gradually change from being small, numerous and scattered to being large, few and clumpy. The authors hypothesize that flavonoid structure, nutrient concentration in the culture medium as well as serum components directly affect MTT reduction by flavonoids in the absence of cells.

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Section: Discussionmentioning

confidence: 99%

Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells

et al. 2006

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“…Oocyte reduction potential was assessed by use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), a water-soluble tetrazolium salt that precipitates as a colored formazan upon reduction. 26 Groups of 25 oocytes were cultured for 4 h in 1.2 mM MTT and then washed several times in phenol-free RPMI medium 1640 (Sigma) supplemented with 0.5% BSA. The oocytes were then transferred into 100 ml of dimethylsulfoxide (DMSO, Sigma) in 96-well plates, and the intensity of the precipitated formazan product was assessed using a plate reader and KC Junior Software (Bio-Tek Instruments, Winooski, VT, USA).…”

Section: Discussionmentioning

confidence: 99%

Genetic variance modifies apoptosis susceptibility in mature oocytes via alterations in DNA repair capacity and mitochondrial ultrastructure

et al. 2006

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Although the identification of specific genes that regulate apoptosis has been a topic of intense study, little is known of the role that background genetic variance plays in modulating cell death. Using germ cells from inbred mouse strains, we found that apoptosis in mature (metaphase II) oocytes is affected by genetic background through at least two different mechanisms. The first, manifested in AKR/J mice, results in genomic instability. This is reflected by numerous DNA double-strand breaks in freshly isolated oocytes, causing a high apoptosis susceptibility and impaired embryonic development following fertilization. Microinjection of Rad51 reduces DNA damage, suppresses apoptosis and improves embryonic development. The second, manifested in FVB mice, results in dramatic dimorphisms in mitochondrial ultrastructure. This is correlated with cytochrome c release and a high apoptosis susceptibility, the latter of which is suppressed by pyruvate treatment, Smac/DIABLO deficiency, or microinjection of 'normal' mitochondria. Therefore, background genetic variance can profoundly affect apoptosis in female germ cells by disrupting both genomic DNA and mitochondrial integrity.

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“…The reaction is attributed mainly to mitochondrial enzymes and electron carriers [38,40,41], although MTT is also reduced by a number of non-mitochondrial enzymes in other cellular compartments [42]. MCN was Fig.…”

Section: Dimebon Enhanced Mtt Reduction In Both Primary Mcn and Diffementioning

confidence: 99%

Dimebon (Latrepirdine) Enhances Mitochondrial Function and Protects Neuronal Cells from Death

Zhang

Hedskog

Petersen

et al. 2010

JAD

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Abstract. Dimebon, a drug currently being evaluated in multiple Phase III Alzheimer's disease trials, has previously been shown to have effects on isolated mitochondria at µM concentrations. Here the effects of nM concentrations of Dimebon on mitochondrial function were investigated both in primary mouse cortical neurons and human neuroblastoma cells (SH-SY5Y). Under non-stress conditions nM concentrations of Dimebon increased succinate dehydrogenase activity (MTT-assay), mitochondrial membrane potential (∆Ψm), and cellular ATP levels. Dimebon treatment had no effect on mitochondria DNA content, implying that mitochondrial biogenesis was not induced. Under stress conditions, mitochondria in Dimebon-treated neurons showed increased resistance to elevated intracellular calcium concentrations, thus, maintaining their ∆Ψm throughout the experiment, in contrast to control neurons, which rapidly lost their ∆Ψm. Moreover, we show that serum-starved differentiated SH-SY5Y cells treated with Dimebon had an increased survival rate as compared to untreated cells. In conclusion, these data demonstrate that Dimebon enhances mitochondrial function both in the absence and presence of stress and Dimebon-treated cells are partially protected to maintain cell viability.

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Mitochondrial and nonmitochondrial reduction of MTT: Interaction of MTT with TMRE, JC‐1, and NAO mitochondrial fluorescent probes

Cited by 263 publications

References 32 publications

Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells

Influence of medium type and serum on MTT reduction by flavonoids in the absence of cells

Genetic variance modifies apoptosis susceptibility in mature oocytes via alterations in DNA repair capacity and mitochondrial ultrastructure

Dimebon (Latrepirdine) Enhances Mitochondrial Function and Protects Neuronal Cells from Death

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