How can I dissociate adherent cells?

Enzymatic or mechanical means can be used to detach the adherent cells from the surface of culture vessel.

Mechanical dissociation is a rapid technique that works well on loosely adherent cells that are sensitive to proteases. Procedures commonly used are shake-off and scraping.

• Shake-off: Some loosely adherent cells and mitotic cells can be dissociated by gentle shaking or rocking of culture vessel. Vigorous pipetting can be employed if shaking is not effective enough.
• Scraping: Cell scraper is sometimes used to detach the tightly adherent cells from the vessel. However, this procedure may damage some cells.

Enzymatic dissociation is generally more time consuming than the mechanical means, which involves the utilization of different proteases. Common examples are trypsin, collagenase and dispase.

• Trypsin: Trypsin is one of the most widely used proteases in cell dissociation, which is applicable for most strongly adherent cells.
• Trypsin and collagenase: This combination of proteases can be used to detach high-density cultures and cultures with multiple layers.
• Dispase: Dispase is used for detaching epidermal cells as confluent, intact sheets from the vessel without dissociating the cells.