I am trying to assess the proliferation of Endothelial cells which are grown in the same dish as a stromal layer. (Can't use inserts to grow endothelial system since for our purposes they need to be together this way)... Now I want to know if anybody had done
a) proliferation assays of endothelial cells in this set up..
b) in our assays we grow endothelial cells for 3 days after adding them to stromal layer..
c) If I consider Brdu assay: how long i need to pulse these cells with Brdu? Followed by BrDu can I separate endothelial cells with let's say anti-CD31 antibody conjugate to Dynal beads?
d) after I separate out these cells with Dynal beads, are these activated? Can I grow these separated cells?
can somebody give me their opinion or suggestion or if they have used this sort of experiment please share your experience..