Worthington Tissue Dissociation Guide
Methods and Materials: Basic Primary Cell Isolation
(Refer to references for application specific parameters)
- For non-perfusion, mince or cut the isolated piece of tissue into 2-4 millimeter pieces with sterile scissors or scalpel.
- Add the tissue pieces to the appropriate buffer or balanced salt solution on ice and wash 2-3 times.
- Add appropriate amount of enzyme(s) and incubate at optimum temperature (usually 37°C) for appropriate time, mixing intermittently.
- Gently disperse the cells by pipeting (trituration).
- Filter the cell suspension through fine mesh.
- Allow the cells to settle and decant excess liquid containing enzymes. Wash and repeat 2-3 times.
- Resuspend cells in appropriate medium or buffer.
- Quantitate cell yield and viability.
- Seed cells for culture, if required.
Perfusion procedures require special equipment and techniques for recirculating the buffers, media and enzymes. Please refer to referenced texts for additional information and guidance.
The Worthington Tissue Tables provide references useful to researchers interested in tissue dissociation and cell harvesting procedures. The references are organized by Tissue and Species type and linked to PubMed citations. The Cell type, Enzymes, and Medium for each reference is provided.
To search by specific criteria, use the Tissue References Search Tool.