I'm working with HUVEC isolated from normal donors but I have some problems. I found a lot of variability in their responsiveness to my stimuli (e.g. TNF-alpha), is it possible to speak about responder and "not-responder" HUVEC according to the different donor? Someone has my same problems? Do you know some principles to discriminate against responder and not responder donors?
Actually, I'm using M199 (Sigma) supplemented with endothelial cell grow supplement and 20%FBS, but I need to eliminate serum for my experiments. Is there someone who has experience in HUVEC culture without serum?
Finally, do you know any continuous HUVEC cell line?
I work with huvec from lonza , it 's an especially medium from Lonza also i used it, but it' with serum. If you stay too much time without serum you will kill them,if you want a continuous huvec cell line, you have to immortalize the line with antigen T. I did it and keep the same markers and maybe you can try to work without serum with immortalized line
I' working with HUVEC using M199 and only 10% of PBS serum without endothelial growth supplement, and yes I've noticed "inter-donors" variability to different stimuli, which is... quite normal, I suppose (i.e. diffent stimuli in course of pregnancy and delivery, different genetic predisposal etc.)
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