If you want to treat cancer cells (i.e. A549 cells) with some stimulus (i.e. LPS) and want to check the effect of any flavonoid, what is the best way for starvation and treatment? Should I pretreat cells with flavonoid ( during starvation time period) and then treat the cells with LPS or should I treat LPS and drug both at the same time?
Heena remove serum once cells are properly attached. wash cells with serum free media. incubate cells for 12-18 h then add your drug.
Dear Hina,
Here is my suggestion base of your treatment.
If you want to see the effect of each condition, you should test each of them separately and together.
For the starvation protocol, I used completed media with 0.01% FBS after reaching the 70% cell confluency in 10%FBS containing media. Like Akash suggested I washed my cells with serum-free media before adding 0.01% serum containing media.
If your purpose to check the effect of LPS and flavanoid stimulus during starvation, when you change the medium for 0.01%, add your treatment parameters separately and together. (you have to do each condition in triplicate)
So, you need test following conditions for a particular time period (12, 24 , 48 hrs)
- control with 10% FBS medium
- control with 0.01% FBS medium
- 10%FBS + LPS
- 0.01% + LPS
-10%FBS + Flavanoid
- 0.01% + Flavanoid
- 10%FBS + LPS + Flavanoid
- 0.01% + LPS + Flavanoid
I hope this helps
Hello
I agree with all suggestions here
look at links , Good Luck
http://articles.mercola.com/sites/articles/archive/2010/06/08/dramatically-effective-new-natural-way-to-starve-cancer-and-obesity.aspx
http://ajpcell.physiology.org/content/301/2/C272.full.pdf+html
https://arxiv.org/pdf/1407.7622.pdf
Dear Hina,
I agreed with Cigdem Ozen .She gives you the best way/protocol for starvation of cancer cells.
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