Hi to everyone,
I'm trying to create a smooth muscle tissue using the fibrin gel obtained mixing fibrinogen and thrombin. For the formation of the tissue first of all I seed smooth muscle cells (500000 cells) on a PDMS substrate (previously treated with laminin 2ug/cm^2) and after 3-4 days I start with the procedure for the creation of the fibrin gel. To this goal I use a combination of 1 ml fibrinogen solution (10mg/ml 0.9% NaCl) and 50 ul thrombin solutions (100 NIH U/ml PBS); I have tried with and without 15 ul CaCl2 solution 20 mM. After that I position the petri dish in incubator for at least 45 minutes and at the end add 1 ml of medium. I have tried this protocol with and without cells. In both cases the fibrin gel forms a layer, the difference is that with cells after at least 2 hours the fibrin gel layer disgregate themself, it is as if it melts; instead without cells the layer remain compacted.
Anyone know why the fibrin gel has this behavior?
Thank you.
try add some aminocaproic acid in media, fibrin gel is easily degradeble
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