Cell culture medium was exposed to powdered polymer (1 100 mg/ml) for a period of 7 days.
The reaction vial kept under 175 psi pressure, stirred at 60°C (temperature monitored by a built-in infrared sensor) and then the medium was exposed to microwave (300 W) radiation for 90s.
The medium was exposed to TCS for 5 hours, then placed back into its original container and stored at 4°C before use.
Two different treatments were used to examine the stability of the S. venezuelae factor: 1) conditioned medium was boiled for 30 minutes, and 2) conditioned medium was exposed to 0.5 mg/mL proteinase K for 1 hr at 65°C prior to heat inactivation of the enzyme at 95°C for 10 minutes.
Later the cell culture medium was exposed to these cured materials or the uncured materials were directly given to the cells [ 29] or were dissolved in cell culture medium [ 30].
This effect was independent of the presence of serum in the culture medium and was only observed when a cell was targeted, but not when only the medium was exposed, confirming that a cell-mediated response is involved.
