I am trying to develop resistance to oxaliplatin in cancer cell lines and want to know if the drug should be given to cells in 5% media or 10% media and if that makes any difference?
Yes, the serum concentration (5% vs. 10%) can make a difference. Using 5% FBS may increase the drug effect because there are fewer proteins to bind the drug, but 10% FBS helps keep the cells healthier during long treatments. I think you should start with 10% FBS to support cell growth and then slowly increase the drug dose to develop resistance. Make sure you maintain the same conditions throughout your experiment to obtain reliable results.
Hello Zainab Saleem
It makes a difference if you use 5% or 10% media. Serum contains albumin which can bind to oxaliplatin forming a complex. This binding can reduce the concentration of free oxaliplatin in the culture media for cellular uptake. Since the active from of oxaliplatin is the unbound form, the binding of oxaliplatin to serum proteins can decrease its cellular uptake. Moreover, lower FBS concentration can better mimic in vivo conditions because cancer cells in the body often experience nutrient limitations and fluctuating conditions, and this may help to lead to a more relevant resistant cell line.
Alternatively, you may use serum-free media supplemented with specific growth factors and hormones to closely mimic the tumor microenvironment. When cells are exposed to a drug in serum-free media, the cells that survive and proliferate are more likely to develop intrinsic resistance mechanisms. This is because the selection pressure would be more direct in the absence of serum’s protective effects. Serum containing media is still commonly used for many cancer cell lines. So, it is up to you to decide whether you would like to go for serum containing media or serum-free media.
You may carry out stepwise drug exposure by gradually increasing the drug concentration over time. This will allow the cells to adapt and evolve resistance mechanisms in response to the drug’s presence.
I have attached a few articles below which may help.
https://star-protocols.cell.com/protocols/4003
Article In vitro Development of Chemotherapy and Targeted Therapy Dr...
https://ar.iiarjournals.org/content/39/12/6443
Best,
Serum's Role in Bioavailability:
Given oxaliplatin's rapid biotransformation and protein binding, FBS might influence its bioavailability and uptake in 2D in vitro systems. You may need to consider serum's effects when designing experiments or interpreting results
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