Trypsin-EDTA is a commonly used solution in cell culture that contains the proteolytic enzyme trypsin and the chelating agent ethylenediaminetetraacetic acid (EDTA). The solution is used to dissociate adherent cells from the surface of a culture dish or flask, allowing them to be harvested for further experiments or subcultured.
Trypsin is a protease enzyme that cleaves peptide bonds in proteins and can break down the proteins that anchor cells to the extracellular matrix. EDTA, on the other hand, is a chelating agent that binds to calcium ions and disrupts the calcium-dependent cell-cell adhesions. When combined, trypsin and EDTA can effectively dissociate cells from the culture vessel.
The optimal concentration and duration of trypsin-EDTA treatment depends on the cell type and the culture conditions. Generally, a brief exposure to trypsin-EDTA, usually 1-5 minutes, is sufficient to dissociate most adherent cells. However, over-exposure to trypsin-EDTA can lead to cell damage or death.
After treatment with trypsin-EDTA, the cells are typically neutralized with a medium containing serum or other cell growth factors to stop the trypsin activity and provide nutrients for the cells. The cells can then be centrifuged and resuspended in fresh medium or used for downstream applications, such as cell counting, flow cytometry, or cell-based assays.