Serum Deprivation - MDA-MB-231 cells - (Dec/18/2008 )
Hi all,
I am working with MDA-MB-231 breast cancer cells and I need to serum deprive them before IGF-1 and insulin treatments. However, they are not well under this condition and I don't want the serum deprivation to have its own effect on my cells. So, I saw some people using Ham's nutrient mixture or McCoy's 5A. Does anyone have any experience/suggestions about this? I would greatly appreciate your comments!!
I am working with MDA-MB-231 breast cancer cells and I need to serum deprive them before IGF-1 and insulin treatments. However, they are not well under this condition and I don't want the serum deprivation to have its own effect on my cells. So, I saw some people using Ham's nutrient mixture or McCoy's 5A. Does anyone have any experience/suggestions about this? I would greatly appreciate your comments!!
Hi!
I am not an expert with these cells and then all the things i say to you are based in my personal experience.
So why you have to deprive your cells with serum??? i think you can easily work with your cells with no serum.
Sometimes i use the lowest concentration of serum (0,2 or 0,5%) for one week and they still well. have u ever tried low concentration of serum??
Regards!
Thank you for your reply!
I need to serum deprive my cells and thus need to use 0% FBS because I need to get rid of all the other background growth factors etc and see the effect of insulin I add. Because the cells are not very healthy in 0% FBS I thought it may be a good idea to use some nutrients other than growth factors. Unfortunatelly, I am new at this and don't have much experience. But thanks anyway!
I need to serum deprive my cells and thus need to use 0% FBS because I need to get rid of all the other background growth factors etc and see the effect of insulin I add. Because the cells are not very healthy in 0% FBS I thought it may be a good idea to use some nutrients other than growth factors. Unfortunatelly, I am new at this and don't have much experience. But thanks anyway!
Hi!!!
mostly cells lines might be damaged with serum deprivation, by the way you are getting rid of the main source of proteins, fatty acids and growth factors.
But How long would u keep these cells with no serum?? if it is for 7-10 days i see no problem. i keep mine when i infect them with West Nile virus (althought i add 0,2% serum but it's almost nothing) for 7 days.
You should try! What are the other nutrtients you told?
Regards!
Reis, V.P.
Hi Reis V.P!
Sorry I couldn't reply earlier. My cell line (MDA-MB-231) does not like the serum starvation (0% FBS) and even after 24 h they start to change and proliferation rate drops. But I need to get rid of all the growth factors present in FBS and therefore I need not to use any FBS. Looking at the papers I saw that people do serum starvation for shorter times like 12 h or 16 h before the actuall treatment and also some people add into serum-free medium some nutrients such as Ham's F12. I just wanted to see whether anybody here has an experience with such conditions.
Thanks alot for your replies!!
Best wishes,
-M-
Hi,
Although I am by no means an expert on this, from time to time we need to serum deprive our cells for a longer time period (several weeks sometimes). We also work with different cells, keratinocytes and fibroblasts. But when we need to deprive our cells, we remove the serum in steps, eg. 2days 5% (normal), 2 days 1% and then to 0%. We supplement the medium, DMEM + HamF12 (ca. 4:1) + some cell dependent additives. You could try this. On the other hand, for short term experiments we starve cells by removing all the serum at once. With added HAMF12 though. You cannot expect cells to feel happy if you remove some essential nutrients at once 
Hope this helps a bit and good luck