An Assay To Evaluate the Anticancer Effects of Lactobacillus Cell-Free Supernatant

Take a multi-well plate containing colorectal cancer spheroids in a growth medium.

The spheroids are three-dimensional aggregates of cancer cells that resemble the physiological tumor microenvironment.

Remove the growth medium and add increasing concentrations of Lactobacillus cell-free supernatant, or LCFS, the culture supernatant obtained from a culture of Lactobacillus, a bacteria of the digestive tract.

LCFS is devoid of bacterial cells but contains active metabolites with anticancer activity secreted by the bacteria.

Bacterial metabolites in the LCFS interact with cancer cells in the spheroid, inducing apoptosis and leading to cell death.

The damage to the cells results in disruption of the spheroid morphology.

Using a microscope, examine the LCFS-treated spheroids to observe an increase in cell death with the increasing concentration of LCFS, indicating the dose-dependent anticancer activity of the bacterial metabolites.