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Collagenase lot selection and purification for adipose tissue digestion
S K Williams1, S McKenney, B E Jarrell
1Department of Surgery, University of Arizona, Tucson 85724, USA.
Cell Transplantation
|May 1, 1995
Summary
Lot variability in crude clostridial collagenase (CCC) affects tissue digestion. Purified collagenase showed reduced efficacy, but combining pure collagenase with trypsin maximized digestion for controlled cell isolation.
Area of Science:
- Biochemistry
- Cell Biology
- Tissue Engineering
Background:
- Crude Clostridial collagenase (CCC) is widely used for tissue digestion in cell isolation.
- Significant lot-to-lot variability in CCC's tissue digestion capacity is a persistent challenge.
- CCC contains multiple enzymatic components beyond specific collagenases and proteases.
Purpose of the Study:
- To evaluate the digestion efficacy of different commercial CCC lots on human adipose tissue.
- To investigate the impact of CCC purification on its tissue digestion capabilities.
- To determine optimal enzyme combinations for consistent cell isolation from adipose tissue.
Main Methods:
- Assessed CCC digestion of human liposuction-derived subcutaneous fat.
- Measured endothelial cell release and adherence post-digestion.
- Purified CCC via dialysis, centrifugation, and liquid chromatography.
- Tested purified CCC alone and in combination with trypsin.
Main Results:
- Observed significant variations in digestion efficacy among different CCC lots.
- Partially purified CCC maintained digestion capacity.
- Completely purified collagenase showed reduced digestion ability.
- Maximum digestion was achieved using completely purified collagenase combined with trypsin.
Conclusions:
- Lot variability in CCC impacts its reliability for tissue digestion.
- Enzyme purification alters collagenase activity, necessitating adjustments for optimal performance.
- Combining purified collagenase with trypsin offers a controlled method for tissue digestion when component identification is critical.