Tissue dissociation/primary cell isolation and cell harvesting are principal applications for enzymes in tissue culture research and cell biology studies. Despite the widespread use of enzymes for these applications over the years, their mechanisms of action in dissociation and harvesting are not well understood. As a result, the choice of one technique over another is often arbitrary and based more on past experience than on an understanding of why the method works and what modifications could lead to even better results.
The goal of a cell isolation procedure is to maximize the yield of functionally viable, dissociated cells. There are many parameters which may affect the outcome of any particular procedure including but not limited to:
1. Type of tissue
2. Species of origin
3. Age of the animal
4. Genetic modification(s) (knockouts, etc.)
5. Dissociation medium used
6. Enzyme(s) used
7. Impurities in any crude enzyme preparation used
8. Concentration(s) of enzyme(s) used
9. Temperature
10. Incubation times
The first four items generally are not a matter of choice. To achieve suitable results the other variable conditions are best defined empirically.
* Text is quoted from the Worthington Enzyme Manual
