Enzyme to Histology contrast in vessel culture - (Jul/01/2010 )
Hello all I am looking for some insight into the enzymatic activity of a vessel excised from an animal over the course of about 20 hours. Specifically it is the aorta of a rat and we double cannulate the vessel and provide suffusate/perfusate to keep inside and outside of vessel adequate nutrients, in hopes of keeping the vessel patent. Solutions are maintained at 37C and CO2 is also provided by an aquarium pump. However I am considering trying a treatment which was quite successful on endothelial cell monolayers on a whole vessel. However the treatment is purported to occur via cAMP stimulation which augments the functions of enzymes and hormones. I believe trying to keep the vessel viable outside the rat is reasonable histologically (Tedgui 1989), but enzymatically I think there is just no chance to have the vessel behave comparatively to the in vivo situation. Can anyone provide some direction or thoughts? I want to get my Ph.D. soon and if I can say the treatment won't work without the enzymes intact, which I think is not possible ex-vivo than I would be nearly finished. Thanks
Is there anything I can clarify to make this problem more answerable?
I think the tissue will remain viable for a while ex vivo, how long is another matter. So long as the tissue is in a nutrient isoosmotic solution perfused with appropriate gas mixtures, then it will behave reasonably normally for a while. I seem to recall people saying that they have kept gut tissue alive for a couple of days in Ringer's solution.
well histologically speaking keeping vessels intact for a week ex vivo has been done. But I don't think that really relays the integrity of the enzymes and the reactions that normally would occur in vivo. This is as far as I have been able to get in terms of predicting what would happen.
basically the main question I want to ask is whether any cAMP related upregulation by treatment would be possible ex-vivo over 20 hours.