Hello Natalia,

I recommend you to use Collagenase D from Clostridium histolyticum (Sigma has one). I have done it many times without problems.

Hopefully this can be helpful.

Best regards,

JP

Dear. Juan Pablo Arab

Hello~ I have a question, too. I am have a big problem in isolating hepatocyte from mouse. what concentration of collagenase D did you apply?

Hello Seungho,

I'm using 35 mg of Collagenase D in 125 mL of the solution with calcium per mouse. Should be at 37 celsius degree ideally.

Let me know if I can help you with anything else,

Juan Pablo

Thank you so much for your kind answer. I solved problem. The problem was...I did pipetting too harsh when resuspending the pellet of hepatocytes in washing buffer after digesting liver and centrifuge. Anyway, I really appreciate for your help.

Hello everyone,

I also have problem in isolating mouse hepatocytes but I am using collagenase type I from Worthington. Could it be that this type has more harsh activity? Does anyone have experience with collagenase type I to recommend a proper concentration? The concentration I have tried is 180U/mL.

Thank you very very much in advance.

Ourania