This study explores the use of folate receptors for targeted delivery of nanoparticles in ovarian cancer cells. By utilizing folic acid-capped nanoparticles, researchers can enhance the selective binding and uptake of these agents in cancer cells.
Folate receptors are membrane-associated glycoproteins overexpressed by a majority of ovarian cancer cells. Exploiting these receptors allows for the targeted delivery of diagnostic nanoparticles into the cancer cells.
Begin with a suspension of the desired folic acid-capped nanoparticle contrast agent conjugates. Add this nanoparticle suspension into an adherent culture of ovarian cancer cells. Incubate the culture for the desired time period.
The folic acid functionalized to the nanoparticles helps recognize the folate receptors expressed on the cancer cells, promoting the selective binding of nanoparticle conjugates with the cells. Subsequently, the plasma membrane invaginates, facilitating the cellular uptake and accumulation of nanoparticles.
Harvest the cancer cells. Next, centrifuge to separate the cells from the supernatant containing any unbound nanoparticles. Discard the supernatant and resuspend the nanoparticle-bearing cells in a suitable buffer for downstream applications.
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.
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