Dear Community,
I am working with 3D cultures. Cells are embedded in fibrin-gel within a scaffold. After my culture I would like to get out as much cells as possible from the scaffold, hence I would like to dissolve the fibrin-gel. Do you have already established protocols? Or could recommend a certain protease? I am thinking about Plasmin, but am afraid that it might be to harsh on the cells since I want to do FACS-analysis afterwards...
Thank you in advance!
Kübrah
Hello Kubrah
Fibrin gels can be dissolved by using 0.05% trypsin-EDTA solution or 100% TrypLE solution. Wash the gel in PBS and then add the mentioned fibrinolytic solution in desired quantity and incubate for 30-40 mins at 37°C. Longer incubation period is recommended for larger quantity of fibrin gel. Collect the dissolved content and centrifuge at low speed. Wash the recovered cells with cold PBS and use for further analysis.
All the best.
search it:
Fibrin gel as a scaffold for photoreceptor cells differentiation from conjunctiva mesenchymal stem cells in retina tissue engineering
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