3T3 Swiss Albino fibroblast cell suspension (Phenol-red free DMEM with 10% FBS and 2 mM L-glutamine) was plated out into 96-well tissue culture plates (1×105 cells/mL, 100 μL/well). The cells were cultured overnight at 37° C., 5% CO2. The extracts and fractions were solubilised in DMSO, and diluted appropriately with media so the extracts were tested at a final concentration of 1000, 10, and 1 μg/mL; 5 μL of sample was added to each well. Cells+samples were incubated (37° C., 3 hours), before the addition of calcium ionophore A23187. Following 20 min incubation, the plate was centrifuged (1000 g, 3 min) and the supernatants were removed. Included on the plate was a positive control (aspirin; 100 μM), and DMSO control, both with and without calcium ionophore A23187.
