This study investigates the anticancer effects of Lactobacillus cell-free supernatant (LCFS) on colorectal cancer spheroids. The research demonstrates how bacterial metabolites in LCFS induce apoptosis in cancer cells, leading to cell death and morphological changes in the spheroids.
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.
For LCFS treatment, serially dilute the prepared thawed LCFS stock solution in fresh growth medium to 25%, 12.5, and 6% concentrations and use a 200-microliter pipette to carefully remove as much supernatant as possible from each well of the plated colorectal cancer cell line cultures.
Then gently add 200 microliters of the LCFS-supplemented growth medium in triplicate to each set of cancer cells, and return the plate to the incubator for an additional 24 to 48 hours.
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