Anti-cancerous Activity of the Rhizome Extract of Alpinia galanga L. Willd. on the Human Liver Carcinoma Cells

Abstract

The rhizome of Alpinia galanga. L. (Willd). was collected from three places i.e, Udupi Shankara narayana temple surrounding area, Shringeri and Madikeri accessions. About 100 gms of the rhizome was taken, dried and powdered to obtain 1 gm of the extract through methanol as the solvent. 1 gm of the solid extract was subjected for filteration through millipore syringe filter., to this about 6ml of cell based assay buffer and 5mg of the MTT buffer was dissolved to obtain a yellow coloured solution. Seed of 100-200μl cell suspension (HepG2-Hepato carcinoma Cell Line) in a 96-well plate at required cell density devoid of the test agent, in the plate in 5% CO2 atmosphere at 24 hrs for required period at 37°C was incubated. After 24 hours the growth medium was removed, freshly prepared each plant extract were serially diluted (100µg serially reduced till 0.78125 µg). Seed 100μl of each dilution in 96-well plate in triplicates after the incubation period, the plates were removed from the incubator and MTT reagent was added to 10% of total volume of final concentration. The volume should be maintained the same as the volume used during the determination optimum cell density and in the process to avoid exposure to light, the plate was wrapped with aluminum foil. The plates were placed back on to the incubator and incubated for 2 to 4 hours. In adherent cells, the culture medium was aspirated without disturbing the monolayer. To the culture volume, the solubilization solution were added in equal amounts. Gentle stirring in gyratory shaker is carried out to enhance dissolution. For complete dissolution, pipetting of the MTT formazone crystals was carried out in dense cultures. Spectrostar Nano ELISA Plate reader was used to read the absorbance at 570nm.

Keywords: Alpinia galanga L. (Willd.), Millipore syringe filter, HepG₂-Hepato carcinoma Cell Line, MTT formazone crystals, Spectrostar Nano ELISA Plate reader

Cite this Article

Naveen Kumar S.P, Maya C. Anti-cancerous Activity of the Rhizome Extract of Alpinia galanga L. Willd. on the Human Liver Carcinoma Cells. Research & Reviews: A Journal of Biotechnology. 2018; 8(2): 1–6p.