Skip to Content
Merck
  • In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

Ecotoxicology and environmental safety (2014-02-18)
Mirella Vazzana, Monica Celi, Cecilia Tramati, Vincenzo Ferrantelli, Vincenzo Arizza, Nicolò Parrinello
ABSTRACT

The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7)M, 10(-5)M, and 10(-3)M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MTT assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells׳ ability to uptake neutral red and viability by MTT assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3)M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Neutral Red, Dye content ≥90 %
Sigma-Aldrich
Neutral Red Solution (0.33%), 3.3 g/L in DPBS, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Neutral Red, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Neutral Red, for microscopy (Bact., Bot., Fl., Hist., Vit.), indicator (pH 6.8-8.0)
Sigma-Aldrich
Neutral Red, certified by the Biological Stain Commission
Sigma-Aldrich
Cadmium, shot, 3 mm, 99.999% trace metals basis
Sigma-Aldrich
Cadmium, granular, 30-80 mesh, ≥99%
Sigma-Aldrich
Cadmium, powder, −100 mesh, 99.5% trace metals basis
Sigma-Aldrich
Neutral Red Solution, Buffered
Sigma-Aldrich
Cadmium, granular, ≥99%, 5-20 mesh
Cadmium, foil, 4mm disks, thickness 0.25mm, as rolled, 99.95%
Cadmium, foil, not light tested, 100x100mm, thickness 0.015mm, 99.7+%
Cadmium, foil, 6mm disks, thickness 0.025mm, 99.7+%
Cadmium, foil, light tested, 25x25mm, thickness 0.025mm, 99.7+%
Cadmium, foil, 8mm disks, thickness 0.005mm, 99.7+%
Cadmium, rod, 150mm, diameter 10.0mm, as drawn, 99.99+%
Cadmium, foil, 8mm disks, thickness 0.25mm, as rolled, 99.95%
Cadmium, foil, 8mm disks, thickness 0.5mm, as rolled, 99.95%
Cadmium, tube, 200mm, outside diameter 2.29mm, inside diameter 1.27mm, wall thickness 0.51mm, 99.95+%
Cadmium, foil, 8mm disks, thickness 0.015mm, 99.7+%
Cadmium, foil, 50mm disks, thickness 0.125mm, as rolled, 99.99+%
Cadmium, rod, 50mm, diameter 10.0mm, 99.999%
Cadmium, foil, not light tested, 25x25mm, thickness 0.002mm, 99.7+%
Cadmium, foil, 8mm disks, thickness 1.0mm, as rolled, 100%
Cadmium, foil, not light tested, 100x100mm, thickness 0.025mm, 99.7+%
Cadmium, foil, 6mm disks, thickness 0.5mm, as rolled, 99.95%
Cadmium, rod, 100mm, diameter 10.0mm, 99.999%
Cadmium, foil, light tested, 50x50mm, thickness 0.015mm, 99.7+%
Cadmium, foil, 8mm disks, thickness 0.01mm, 99.7+%
Cadmium, foil, not light tested, 50x50mm, thickness 0.005mm, 99.7+%