We want to use thoracic aorta isolated from rats yesterday then we preserved in a refrigerator then we used in tissue bath after that we pre-contracted aorta by PE then relaxing by ACh.
It depends what you want to use it for. The muscle should be ok, not so sure about the endothelium. If you want to isolate protein, fine, for RNA I would use a fresh aorta.
If sterility has been maintained and you are seeking to isolate endothelial and/or vascular smooth muscle cells for in vitro expansion, you might still be able... it is worth a try!
Dear Katharina Mahn
Dear Ira M. Herman
I want to use in isolated tissue preparation
We bind this aorta between two stainless steel and connected with force transducer to determination contraction and relaxation
Are OK or not
Thanks for you reply
In which case I would use a fresh tissue if you need endothelium-dependent relaxation (ACh) etc. If you are looking at endothelium-independent relaxation it would be fine to use. But why not give it a try, a one-off dose of ACh will tell you quickly if the endothelium is still ok.
Thank you Katharina Mahn
Depending on your answer we tried to do
Thank you very much
I will not advise to use this tissue. You can use fresh prepared tissue up to 4 hours after preparation, provided that you keep it in relatively cool and aerated (carbogen) physiological solution. When kept in the refrigerator, smooth muscle cells may loose their sensitivity. Moreover, cells may produce large amount of reactive oxygen species due to keeping conditions and mostly to re-oxygenation, that may drastically affect endothelial cells and smooth muscle cells as well.
Based on our previous experience, it will work, but may not yield optimal results compared to fresh preparation. Therefore, I would prefer freshly isolated preparation for these type of experiments.
Thank you Télesphore Benoit Benoît Nguelefack
Thank you Laxmansa C Katwa
I agree with Telesphore. You can not use aorta for ex vivo experiments after it was stored over night at 4C. You should plan your experiments carefully and be able to use the specimen immediately after the dissection. After dissection you should keep your specimen in oxygenated Ringer solution. It should be completely immersed in solution and preferably pinned down for optimal stretch, prior to the experiments. In such solutions you can keep the aorta only for the few hours still not over night.
If the main purpose of the experiments is to study sample contraction and relaxation, in vitro, but in conditions not too far from physiological ones, it seems advisable to work with a fresh sample, as was argued by the researchers that gave answers above.
HI, I've just seen your question. I think you can also consider these two (old) papers: Shibata S., Circ Res 1969,24:179-187 and McIntyre CA et al., Br J Pharmacol 1998,123:1555-60
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