Author/Authors :
Abnosi M، Mohammad Hussein نويسنده Department of Biology, Arak University of Medical Sciences, Arak, Iran , , Jafari Yazdi، Zahra نويسنده Department of Biology, Arak University of Medical Sciences, Arak, Iran ,
Abstract :
Objective: Although arsenite is toxic it is currently recommended for the treatment of malignancies.
In
this
study
the effects
of
sub-micromolar
concentrations
of sodium
arsenite on
the viability,
morphology
and
mechanism of cell
death of rat
bone marrow mesenchymal
stem cells (BMCs) over 21 days was investigated.
Materials and Methods: In this experimental study, BMCs were extracted in Dulbecco’s Modified Eagles Medium (DMEM) containing 15% of fetal bovine serum (FBS) and expanded
till the 3
passage. The cells were treated with 1, 10, 25, 50, 75 and 100 nM of sodium
arsenite
for
21
days
and
the
viability
of
the
cells
estimated
using
3-(4,
5-dimethylthiazol-2-yl)-2,
5
diphenyl
tetrazolium
(MTT)
and
trypan
blue
staining.
Cells
were
then
treated
with
the
selected
dose
(25
nM)
of
sodium
arsenite
to
determine
their
colony
forming
ability
(CFA)
and population
doubling
number
(PDN). Morphology of
the cells was
studied using
florescent
dyes,
and
the
integrity
of
the
DNA
was
investigated
using
the
comet
assay
and
agarose
gel
electrophoresis.
The
terminal
deoxynucleotidyl
transferase
dUTP
nick
end
labeling
(TUNEL)
and
the
caspase
3
assay
were
then
applied
to
understand
the
mechanism
rd
of cell death. Data was analyzed using one way ANOVA, Tukey test.
Results: A significant reduction of viability, PDN and CFA was found following treat-
ment of BMCs with 25 nM sodium arsenite (p < 0.05). Cytoplasm shrinkage and a significant decrease in the diameter of the nuclei were also seen. Comet assay and agarose gel electrophoresis revealed DNA breakage, while positive TUNEL and activated
caspase 3 confirmed the apoptosis.
Conclusion: A low concentration of sodium arsenite (25 nM) caused reduction of viability due to induction of apoptosis. Therefore, long term exposure to low dose of this chemical may have unwanted effects on BMCs.
