In-vitro Cytotoxic Activity of E. coli Outer Membrane Vesicles (OMVs) Against Breast Cancer (MCF-7) Cell Line
Keywords:Cytotoxic Activity; E. coli; outer Membrane Vesicles (OMVs); breast cancer (MCF-7).
Twenty-five samples of urine were collected from patients suffering Urinary Tract Infection (UTI) from
hospital in Baghdad,twenty-two bacterial isolated (88%) were identified. From our result showed the E. coli
number 5 is the best bacterial virulence factor for producing OMV, Tumor cells (1x104 – 1x106 cells/ml)
have been grown in 96 flat well micro-title plates, with a final volume of 200 cells per well. The microplate
was gently shackled and covered by sterilized parafilm then incubation of the plates at 37C°, 5 % CO2 for
24hrs. Two folded serial dilutions of the desired compound (12.5, 25, 50, 100, 200, 400µ g/mL) were added
to the wells after incubation. Triplicates were used per each concentration as well as controls (serum-free
medium treated cells). Plates were incubated for specified exposure time (24 hours) at 37C°, 5 percent
CO2. Applied 10µ liters of MTT solution to each well after exposure. The absorbance was determined by
a wavelength of 575 nm using an ELISA reader. Statistical analysis was performed on the optical density
data to calculate the compound concentration required to cause a 50 percent reduction in cell viability for
each cell line. The rate of cell growth inhibition (the cytotoxicity percentage) was calculated and plot of
% cytotoxicity against sample concentrations assisted to calculate the concentration which exhibited 50%
cytotoxicity (IC50). Calculate: cell number: determine cells per milliliter.
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