Endothelial permeability - looking for help with my project - (Feb/15/2012 )
I have started my PhD project one year ago in the field of matrix biology. It still doesn't really have a research question, however, I am supposed to work on endothelial permeability.
I wanted to have an in vitro system to test the effect of a substance X we have purified in the lab on the permeability of endothelial cell monolayers. Unfortunately, the test (which is done in membrane inserts for 24 well plates and uses a fluorescently labeled dextran as tracer) didn't work. Which in this case means that I couldn't increase the permeability with any substance I tested (e.g. histamine, VEGF, EDTA, bradykinin). So after one year I still don't have any results and I need to think about a change in the project. My supervisor is not really helpful, he only suggests further reading (which I have done but which doesn't really help me having ideas). I already started working with mice and looking at the effect of substance X on tumor vascularure.
I really need some help/input here, because my funding ends on the 30th April and I will only get more money if I have results and a project plan for the next year. At the moment I am thinking of just giving up because I am so frustrated...
Any suggestions are most welcome!
Attached is an interesting paper related to monolayer permeability. They used texas red-labeled dextran but likely had a different set up. You can also measure TEER. All the info and methods is in the link below.
Look up trypan blue permeability assay or how you could use permeability chambers for performing endothelial cell permeability assay using albumin texas red. Hope this helps!