The A549 non-small cell lung adenoc

The A549 non-small cell lung adenocarcinoma cell line, SK-Hep-
1 hepatocellular carcinoma cell line and MCF-7 human breast adenocarcinoma
cell line from ATCC were used for the cytotoxicity
test. The cells were grown in RPMI 1640 (A549 and SK-Hep-1), or
DMEM (MCF-7) supplemented with 10% fetal bovine serum (FBS),
100 U/ml of penicillin, 100 lg/ml of streptomycin and 125 ng/ml
of amphotericin B. All cultures were maintained at 37 C in a
humidified atmosphere of 5% CO2.
Cytotoxicity assays of crude polysaccharides from fresh fruit
bodies and dried fruit bodies, on SK-Hep1 and MCF-7, were evaluated
in vitro using the MTT assay. Briefly, cell suspensions in culture
medium were seeded in 96-well plates (1  104 cells/well),
and incubated at 37 C in a humidified atmosphere of 5% CO2. After
24 h, additional medium (100 ll) containing test sample was
added to each well, followed by further incubation for 48 h. Then,
the wells were replaced and incubated with fresh culture media
containing MTT (0.5 mg/ml) for 2 h at 37 C. Finally, the media
were removed and DMSO was added to the wells (100 ll/well),
and absorbance was measured at 550 nm in a microtitre plate
reader and subtracted from the absorbance at 650 nm. The number
of viable cells was determined from the absorbance. Assays were
performed in quadruplet wells. Data were expressed as percent
viability compared with control (mean ± SD).