Trypsinisation

Trypsinisation

Theory

Trypsin is an enzyme isolated from the pancreas of animals. It is one of the least-specific of the serine proteases, so cuts peptide chains regularly. When applied to tissue it disrupts the proteinaceous components of the extracellular space, leaving the intracellular proteins (within the cell membrane) unharmed. This frees cells from the extracellular tissue, aiding some types of microscopy.

Practice

1. Samples of tissue were incubated in Hank's Buffered Saline Solution (HBSS) with a 2.5% trypsin solution for 30 minutes with gentle agitation of the tissue.
2. The supernatants were then gently removed by pipette and transferred to a clean centrifuge tube. Dispersed cells were pelleted at 3000g for 10 minutes; the supernatant was discarded.
3. The pelleted cells were resuspended in a little HBSS, sandwiched between a slide and coverslip then viewed.

Safety

MSDS data sheets for trypsin indicate that it is harmful if swallowed, may act as a sensitiser by skin contact and is a skin, eye and respiratory irritant. Appropriate measures should therefore be taken.