Quantitative reduction of methyl tetrazolium by fresh vein homograft biopsies in vitro is an index of viability

B. Galambos, L. Csönge, A. Olah, R. Von Versen, L. Tamas, P. Zsoldos

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Objective: The color density of the methyl tetrazolium (MTT) test is proportional to mitochondrial enzyme activity thus reflecting cellular viability. The aim was to evaluate the MTT test as a viability assay for vein homograft studies. Materials and Methods: Fresh intact vein samples were harvested during multi-organ procurement. The reliability of the MTT assay was tested by a fluorescent dye combination (1 μg/ml propidium iodide PI and 4 μM/ml SYTO-16 stains). The enzyme kinetics of the reaction was also investigated. The optimal reagent concentration, biopsy size and incubation period were established. Results: There was a linear relationship between the vein homograft's weight and the pigment production activity. A nonspecific reaction (8.6%) was observed in negative controls. The MTT cleavage up to 0.1% (w/v) follows the Michaelis kinetics. The Michaelis constant (2,805 ± 130 μM), the maximal velocity (196 ± 2.2 × 10-5 μM s-1) and the velocity constant (6.98 ± 0.2 × 10 -7 s-1) was calculated. The viability assessed by fluorescent dyes simultaneously visualized the live/dead cell ratio, which can be calculated by image analysis software. Conclusion: The use of MTT in colorimetric assays offers high sensitivity. The assay is simple, inexpensive, and reproducible in vein homograft studies.

Original languageEnglish
Pages (from-to)371-375
Number of pages5
JournalEuropean Surgical Research
Volume36
Issue number6
DOIs
Publication statusPublished - Dec 31 2004

Keywords

  • Fluorescent dye
  • Homograft
  • MTT test
  • Viability assay

ASJC Scopus subject areas

  • Surgery

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