Quantitative Reduction of MTT by Hearts Biopsies In Vitro is an Index of Viability

Ferrera, René; Larese, A.; Berthod, F; Guidollet, J; Rodríguez, C.; Dureau, G; Dittmar, A.

doi:10.1006/jmcc.1993.1121

Cited by 28 publications

(23 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In living cells, mitochondrial reductases convert the watersoluble, yellow tetrazolium salt (MTT) into a water-insoluble formazan precipitate. 25 The production of formazan was linearly related to the amount of myocardial tissue in each assay ( Figure 1A). As shown in Figure 1B, formazan production was stable over the first 24 hours and remained Ͼ90% of starting levels between days 0 and 3 of culture, indicating preserved metabolic activity and viability for Ն72 hours.…”

Section: Myocardial Strip Viabilitymentioning

confidence: 97%

See 1 more Smart Citation

Bucindolol Displays Intrinsic Sympathomimetic Activity in Human Myocardium

et al. 2002

View full text Add to dashboard Cite

Background-Most clinical studies have shown that ␤-adrenergic receptor antagonists improve long-term survival in heart failure patients. Bucindolol, a nonselective ␤-receptor blocker, however, failed to reduce heart failure mortality in a recent large clinical trial. The reasons for this failure are not known. Bucindolol has partial agonist properties in rat myocardium, but whether it has agonist activity in human heart is controversial. To address this, we measured the ability of bucindolol to increase cAMP accumulation in human myocardium. Methods and Results-Myocardial strips (Ϸ1 mm3 ) obtained from rat and nonfailing human hearts were confirmed to be viable for Ն48 hours in normoxic tissue culture by MTT assay and histology. Freshly isolated strips were exposed to ␤-adrenergic antagonists and agonists and assayed for cAMP. In both rat and human strips, the full ␤-adrenergic agonist isoproterenol raised cAMP levels by Ͼ2.5-fold at 15 minutes. Carvedilol and propranolol had no effect on basal cAMP levels, whereas metoprolol reduced basal cAMP by Ϸ25%. In contrast, bucindolol and xamoterol increased cAMP levels in a concentration-dependent manner in both rat and human myocardium (maximum 1.64Ϯ0.25-fold and 2.00Ϯ0.27-fold over control, respectively, PϽ0.01 for human tissue). Conclusions-Bucindolol exhibits Ϸ60% of the ␤-adrenergic agonist activity of xamoterol in normal human myocardial tissue.

show abstract

Section: Myocardial Strip Viabilitymentioning

confidence: 97%

“…The strips were then washed with PBS, and 1 mL 100% DMSO was added for 50 minutes with vigorous shaking to dissolve the resulting formazan crystals. 25 Color production was measured by spectrophotometry at 560 nm.…”

Section: Mtt Assaymentioning

confidence: 99%

Bucindolol Displays Intrinsic Sympathomimetic Activity in Human Myocardium

et al. 2002

View full text Add to dashboard Cite

show abstract

“…The plate was then filled with DMEM with antibiotics and placed overnight at À208C and 8 h at 48C. This procedure was repeated three times and the exhaustive elimination of living cells was assessed by MTT staining as described previously (Ferrera et al, 1993). After tissue devitalization, 2.1 Â 10 5 HUVEC/cm 2 were seeded on the sponge.…”

Section: Reconstructed-connective Tissue Devitalizationmentioning

confidence: 99%

Extracellular matrix deposition by fibroblasts is necessary to promote capillary‐like tube formation in vitro

Berthod

Germain

Tremblay

et al. 2006

Journal Cellular Physiology

126

View full text Add to dashboard Cite

The contribution of the cellular and fibrillar microenvironment to angiogenesis still remains unclear. Our purpose was to evaluate the effect of the extracellular matrix deposited by fibroblasts on the capacity of human endothelial cells to form capillaries in vitro. We have drastically decreased the amount of extracellular matrix surrounding fibroblasts in our model of endothelialized-reconstructed connective tissue (ERCT) by culturing it without ascorbate. Under these conditions, the number of capillary-like tubes (CLT) formed by endothelial cells was reduced by up to 10-fold after 31 days of culture compared to controls. This decrease was due neither to a variation of MMP-2 and MMP-9 secretion, nor to a reduction in the number of fibroblasts and/or endothelial cells, or a diminution of fibroblast growth factor 2 (FGF2) synthesis. The secretion of vascular endothelial growth factor (VEGF) by fibroblasts accounted for 25-70% of the capillary-like tube formation when tissues were cultured in the presence or absence of ascorbate, as demonstrated by VEGF-blocking studies. The culture of endothelial cells on a similar extracellular matrix but in the absence of living fibroblasts did not promote the formation of CLT, even when tissues were fed with fibroblast-conditioned medium. Thus, the deposition of a rich extracellular matrix by living fibroblasts appeared necessary, but not sufficient to promote capillary-like formation. Fibroblasts seem to induce endothelial cells to spontaneously form CLT by secreting and organizing an abundant extracellular matrix, which creates a microenvironment around cells that could in turn trap growth factors produced by fibroblasts and promote three-dimensional cell organization.

show abstract

“…The succinate dehydrogenase was proposed as the enzyme responsible for the tetrazolium salt reduction [1], but in fact a group of enzymes and coenzymes (NAD, NADP) are involved [2]. We have previously shown that tetrazolium reductase activity may be used as an index of viability of fresh and stored heart biopsies [3]. TTC staining is also a widely accepted technique for the assessment of infarct size and delineation of tissue necrosis.…”

Section: Introductionmentioning

confidence: 99%