Good day to all, I am currently working with AGS cells for my MTT assay and would like to take the absorbance readings after 24h, 48h and 72h of treating the cells with certain plant extracts. Most of the papers I referred to fixed the cell number to 5000 - 10000 cells/well for these three time points. Can anyone advice if there is a need to reduce the cell number for longer incubation period if taking into consideration the ongoing cell proliferation of the surviving cells.
In general, no, you have to seed at the same concentration to keep your variables as low as possible.
If you are concerned about overgrowth (and this depends on the cell you are using, and whether they are differentiated or not, and some other stuff), you can lower the % of FBS.
Agreed - I've never seen someone seeding at different starting densities to account for different timepoints. If you're worried, I'd seed at the low end of what you usually use for passaging your cells. I like Shen-An's idea of reducing FBS.
If your assay is intended to measure cytotoxic or anti-proliferative effects of the plant extracts, you could also move all your timepoints up if your cells are growing quickly. Fast-growing cells mean any differential effect should show up faster.
I see. Thank you very much for the responses, much appreciated. It is interesting to know that fast growing cells will show the effect faster...something for me to explore further.
- Badges
- Science topic
- Similar topics
- Papers