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Cytotoxicity against PANC-1 cell
Cytotoxicity against PANC-1 cell

Exploratory Research

Measured value Ratio to control (%)
Animal species Human
Indication of the result The ratio (%) of viable cell number under test drug treatment to that for the control condition (autoclaved water for crude drug extract or DMSO for compound) is shown. Values less than 100 mean cytotoxic effect.
Target diseases of assay Cancer
Assay level Cell-based
Protocol of assay 1) Each crude drug extract was passed through 0.22 μm filters for sterilization. Each compound which was extracted from the herbal medicine (10 mM, 2 μl) was mixed with 6 μL of phosphate-buffered saline (PBS). 2) PANC-1 cells were seeded in 96-well micro plates at a density of 1500 cells/well (4.8×10^4 cells/cm^2) or 3000 cells/well (9.6×10^4 cells/cm^2) and cultured for 24 h at 37 °C in the D-MEM containing 4.5 g/L glucose and supplemented with 10% FCS, 2 mM glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin. 3) The medium was removed and the cells were washed with PBS three times. The new medium (97 μL per well for crude drug and 98 μL per well for compound) was added. 4) Each test drug solution (3.0 μL for crude drug and 2.0 μL for compound) was added to the medium, and the cells were incubated for 24 h at 37 °C. 5) The medium was removed and the cells were washed with PBS three times. The new medium (100 μL per well) was added. 6) 10 μL of WST-8 reagent was added and the cells were incubated for 3 h at 37 °C. 7) The viable cell number in each well was measured by the difference of the absorbance at 450 nm and 650 nm using a microplate reader.
Remarks  
【Results of bioassay】
Display method Crude drug Kampo formula Compound
Crude drug-Compound
Name of experimental material
(compound/crude drug/Kampo formula)

 

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Division of Pharmacognosy,
Department of Medicinal Resources,
Institute of Natural Medicine,
University of Toyama
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