2.5.1. Evaluation of the Chromosomal aberrations (CAs) and the
Mitotic Index (MI)
BM cells were prepared according to the colchicine-hypotonic citrate
technique for CAs. Briefly, 2 hours before sacrifice, rats were
IP-injected with colchicine (2mg/kg BW); the femur bone of each animal
was taken and cleaned from tissues and muscles, and then the femoral
marrow was flushed out using (0.075M) potassium chloride into a
centrifuge tube and centrifuged at 2000rpm for 10minutes. Additionally,
the cells pellets were fixed in (1:3) acetic acid/methanol (v/v), which
was repeated three times; then the cell suspension was dropped on coded
and sterile cleaned frosted slides and then dried followed by staining
with 10% Giemsa stain; and finally, the slides were examined under a
light microscope (Japan, Meiji). The frequency of CAs was scored in at
least 100 meta-phase plates per animal; moreover, the chromatid -gaps
and -break chromosome gaps and breaks, ring, deletion and exchanges were
recorded; and the MI was obtained by counting at least 1000 cells per
animal for dividing cells (18) (19).