Functionalized Nanoparticle Targeting of Ovarian Cancer: A Technique to Facilitate Folic Acid-conjugated Nanoparticle Contrast Agent Uptake by Ovarian Cancer Cells

First, mix the folic acid-capped copper sulfide nanoparticles at the proper concentration in fresh RPMI media. Add this nanoparticle solution to each well of the prepared 24-well plate that contains SKOV-3 cells at a concentration of 400 micrograms per milliliter. Incubate at 37 degrees Celsius with 5% carbon dioxide for two hours.

After this, trypsinize the cells with 0.5 milliliters of 0.25% trypsin and EDTA. To neutralize the trypsin, add at least 1 milliliter of folic acid-free RPMI 1640 complete growth media and centrifuge the cells at 123 x g for six minutes. To wash the cells, remove the supernatant.

Resuspend the cells in 2 milliliters of PBS and centrifuge at 123 x g for six minutes. Repeat this wash step twice to remove any unbound nanoparticles.

Then, resuspend the cells with 1 to 2 milliliters of a 2% Tween solution in PBS. Count the cells using a hemocytometer and Trypan Blue. Dilute the cells in a solution of 2% Tween and PBS to the chosen concentration for detection.