Enzymatic Dissociation

1. Under the laminar flow hood, place your cell culture flask with your adherent cell line. Remove your medium being careful not to disrupt the cell layer. Wash your cells with warm washing solution (e.g. DPBS) by carefully pipetting it to the culture flask and gently whisking the cell culture flask in an “8” motion. Discard the washing solution.

2. Add the disassociation agent and gently rock the flask to cover all the surface. Incubate the cells for 2-3 minutes, use our timer not to lose track of time. Note that the time for the digestion is greatly dependent from cell type and disassociation agent.

3. After the incubation time retrieve the cells and check under the light microscope for detachment. You can facilitate the process by carefully tapping the culture vessel. Once the cells are detached (they will appear rounder under the microscope), halt the action of the dissociating agent by adding pre-warmed cell culture medium. Resuspend the cells and collect them in a falcon tube.

Mechanical Dissociation

For cell lines susceptible to proteases, a mechanical dissociation method may be more suitable. We offer a broad range of scrapers to meet your needs.

Prepare the cells for seeding, freezing or other cell-based analysis by counting them.