I want to solubilize decellularized ECM particles in order to get a homogeneous solution.
As I have seen in the literature, a common way to digest dECM is using pepsin/HCl. To do so, I used Hcl 0.01 M and 10 mg/mL dECM in 1 mg/mL pepsin at 25 C . once again the Hcl, dECM and pepsin respectively in proportion1,5,2(0.2 ml, 1mg, 0.4mg) were used (at 37 C).But I didn't get the desired result and the particles of the matter remain insoluble.
One reason could be that the pH is not in the right range for pepsin activity. Pepsin activity is highest at 2. Once you add the HCl to the dECM please check the pH and adjust it to 2.
Also leftover decellularizing agents also interferes with the pepsin digestion. Make sure the decellularized tissue is washed extensively to remove those.
Also the ECM should be milled very fine. It is difficult to digest very large particles.
Hopefully these should help
Hi,
What is the originate of your dECM?
1. Check pH and adjust to around 2 as Kumar Chokalingam suggested.
2. Perform the digestion at 37oC with agitation.
Good luck!
The pH was 2, but I will be more careful about washing in the new series. Thank you very much for your tips.
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