2.5. Cell Culture, Stimulation of RAW 264.7 Cells with LPS and Bacteria lysate

The Pondicherry Center for Biological Science and Educational Trust gifted the RAW 264.7 cell line. At a temperature of 37 °C and a carbon dioxide concentration of 5%, the RAW cells were incubated with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 μg/mL penicillin. The probiotic cell-free lysate from antimicrobial activity was dissolved in the DMEM medium used for the culturing. In a Petri dish of 10 cm in diameter, the RAW cells were cultivated and then subjected to one of the following treatments: control group, LPS treatment group for 30 min, and LPS+ probiotic lysate (12.5 μg/mL, 25 μg/mL, and 50 μg/mL) group. After the cell culture was finished, the cells from the various treatment groups were collected for assessment of cell viability and cytokine estimation.