Worthington Tissue Dissociation Guide

(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.