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.
Next: Equilibration with 95%O2:5%CO2
Tissue Tables (references, grouped by tissue type and species)
Note: We have not limited the references listed to only those papers
using Worthington enzymes. Generally speaking, the tissue dissociation enzymes
offered by Worthington can be used interchangeably for most preparations