Exposure of Caco-2 Cells to Pesticide Metabolites from Pesticide-Treated Carrot Callus for Cytotoxicity Study

Proceed to perform the exposure test. Once enough cells are harvested for the same, gently tap the flask wall containing the cells to distribute them uniformly and transfer the cell suspensions to a 15 milliliter centrifuge tube. Dilute the cell suspension with DMEM in combination with cytometry to achieve the desired cell density.

Gently tap the flask wall to ensure uniform distribution of cells. Next, add 100 microliters of PBS to the outer wells of the 96 well plate to prevent the culture medium from evaporating due to the edge effect. Add 100 microliters of the cell suspension to the left wells of the plate and allow it to rest for 10 minutes.

Incubate the cells at 37 degrees Celsius with 5%carbon dioxide for 48 hours. After removing the 96 well plate from the incubator, discard the culture medium. To set up a pesticide metabolite group, add 100 microliters of the different prepared pesticide residue exposure solutions to each designated well.

Then add 100 microliters of the different prepared parent pesticide exposure solutions to each designated well for comparison to establish a pesticide parent group. For setting up a blank control, add 100 microliters of DMEM containing 0.3%DMSO to each designated well. Incubate the cells at 37 degrees Celsius with 5%carbon dioxide for 24 hours.