I would like to know how collagenase is used to digest adipose tissue. I've only ever seen it sold in powder form. Does this powder have to be diluted or mixed with something? Or is the powder just applied to the adipose tissue?
Hi Ellie,
You can dissolve the powder in DMEM or F12 media. Could you please specify the purpose?
Just for isolating adipocytes or preadipocytes from adipose tissue. I don't understand how the collagenase is applied to the adipose tissue?
We dissolved collagenase and dispase in media or PBS just before it was used on minced, breast tissue. I do not remember the concentration we used--but it is published. The lipid in breast tissue does interfere with the enzyme activity, so you must first separate adipose cells from the lipid. I think your adipose cells would get lysed while doing this. SO, perhaps you need a different (Non-enzymatic) method to isolate your adipose cells?? Since the lipid would make adipose cells “light ”, differential centrifugation may work. Hope this helps!
Dear Ellie,
Usually collagenase is dissolved in either serum free media,PBS or HBSS at a concentration of 0.1 to 0.2% for efficient digestion of the adipose tissue.It completely depends on what you are trying to obtain,adipocytes,preadipocytes or stomal vascular fraction containing stromal stem cells and endothelial cells.You should standardize the digestion time for fat obtained from different regions of the body as Dr.Venil mentioned that the lipid content could be an interference in digestion..
Regards
Charan
You could mince the adipose tissue in a petri dish placed on ice and then transfer it to a 50ml tube. prepare 10ml of collagenase type 1 solution pr 3g of tissue by dissolving 2mg/ml collagenase in serum free media and filter a 0.2um filter. you then incubate at 37 degrees preferably with gentle shaking
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