I have prepared thymoquinone alginate beads, and I want to investigate the cytotoxicity of my product against colon cancer cell.
Is there a recommended way to do that:
placing the beads in the 96 well plate directly or extracting thymoquinone from the beads first then applying on the cells
There are a few options for testing the cytotoxicity of thymoquinone alginate beads against colon cancer cells:
1. Extract thymoquinone from the beads first, then apply the thymoquinone to the cells. This allows you to know the exact concentration of thymoquinone the cells are exposed to.
- Extract thymoquinone using a solvent like ethanol or methanol. Sonicate or vortex the beads to release the thymoquinone.
- Evaporate the solvent to collect the thymoquinone. Dissolve in DMSO or culture medium for cell treatment.
- Treat cancer cells (eg. HCT116, HT29) with thymoquinone at different concentrations (10 μM - 100 μM). Assess cell viability after 24-48 hrs.
2. Directly add the thymoquinone beads to cells:
- Place beads in 96-well plate, seed cells on top or around beads. The thymoquinone will diffuse out.
- This makes it harder to control the exact dose, but may better mimic delivery in vivo.
- Assess cell viability after 24-48 hrs. Compare to control beads without thymoquinone.
3. Extract media conditioned with beads, apply to cells:
- Incubate beads in media for 24 hrs, collect conditioned media.
- Apply conditioned media to cells. This will contain released thymoquinone.
- Assess cell viability after 24-48 hrs compared to unconditioned media.
So, extracting thymoquinone first allows better dose control, while direct bead addition mimics in vivo delivery.
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