I am going to perform a small study measuring responses to adipokines and have only a small funding pot allocated.
Much of the original work with HUVECs was done in M199 supplemented with 20% FBS and 2 mM glutamine plus penn/strep. This was the original formulation of Jaffe in the seminal paper linked below:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC302542/pdf/jcinvest00186-0087.pdf
Since then a number of improved formulations have been used, adding various growth factors or reducing serum content.
If cost is your main driver, then the above media is going to be most cost effective and has a long track record.
However, your adipokines may be completely swamped by the presence of 20% serum, so you may end up needing to use a low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media:
http://www.lifelinecelltech.biz/category_s/161.htm
Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
Alternately, if your adipokine treatments are relatively short (say 12 hours or less), you can just use the M199 formulation without serum, but be sure to add 2% BSA so there is some protein. This remains a common way to induce quiescence in the monolayer.
Much of the original work with HUVECs was done in M199 supplemented with 20% FBS and 2 mM glutamine plus penn/strep. This was the original formulation of Jaffe in the seminal paper linked below:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC302542/pdf/jcinvest00186-0087.pdf
Since then a number of improved formulations have been used, adding various growth factors or reducing serum content.
If cost is your main driver, then the above media is going to be most cost effective and has a long track record.
However, your adipokines may be completely swamped by the presence of 20% serum, so you may end up needing to use a low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media:
http://www.lifelinecelltech.biz/category_s/161.htm
Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
Alternately, if your adipokine treatments are relatively short (say 12 hours or less), you can just use the M199 formulation without serum, but be sure to add 2% BSA so there is some protein. This remains a common way to induce quiescence in the monolayer.
Hi James.
Personally I've had best results with EGM-2 medium from Lonza, 500ml. It is not so cheap, but is very easy to get and allows you to obtain primary cell cultures in optimal growth conditions. Alternatively, you can buy growth factors for endothelial cells (such as hEGF, VEGF, hFGF-B and r3-IGF-1) and integrate them in a basal medium with FBS and gentamicin/amphotericin B.
Hi James,
for our HUVECs we use ENDOPAN3, unfortunately it's not quite cheap, but the cells are growing really good. There must some growth factors and antibiotics added.
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