A common procedure used to isolate primary cells is to dissociate the tissue with a bacterial enzymatic mixture containing collagenase. The most commonly used reagent is “crude” collagenase, an enzyme mixture minimally purified from Clostridium histolyticum culture supernatants that contains collagenase and other proteases¹. The lot to lot variability in the enzyme composition and performance in the customer’s application directly reflects the biological variability of the culture conditions. Most customers “pre-qualify” lots by testing a sample from a specific lot in their cell isolation application prior to purchase of additional product. This post discusses a method to reveal the collagenase profile found in lots of crude collagenase used successfully in cell isolation procedures with the goal of developing protocols for these applications using purified collagenases.

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