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Materials and Methods:
Juices – Organic cranberry juice and Montmorency cherry juice were obtained. ( refer to appendix a). The pHs of the juices were measured. Four different modifications were made to the juices to produce sterilized juice, neutralized Juice, sterilized concentrated juice and sterilized neutralized concentrated juice. Various concentrations of the juices were made with HT29 cell media (CDMEM). The pHs of the juices were measured. Four different modifications were made to the juices to produce sterilized juice, neutralized Juice, sterilized concentrated juice and sterilized neutralized concentrated juice.
Epithelial cell cultures – A lining of HT29 epithelial cell cultures were grown in plastic wells. The cell cultures were exposed to various concentrations of sterilized neutralized cranberry and cherry juice overnight in an incubator at 37°C. After removing the supernatants from the wells, the fluorescence of living cells in each plate exposed to concentrations of juices were then compared to a control by adding 25 μl of calcein-AM to each well and then using the FCA machine.
IL-8 Production by HT29 cells – An IL-8 Enzyme-linked Immunosorbent Assay (ELISA assay) was done with the supernatants diluted 1:10 from the cell cultures to determine how much IL-8 HT29 cells made when cells were exposed to different concentrations of fruit juice. (refer to appendix b for IL-8 ELISA procedure). Further dilutions of the supernatants were made for IL-8 readings that were above the standard curve (200 pg). |