in cell culture .we use trypsin/EDTA to remove the adhesion of cells from plate. how positive charge removed so cells become suspended and any other chemical can be used to make the same function ?
Andrew Octavian Sasmita
Trypsin is a serine protease which cleaves amino acids, such as lysine arginine, which are essential in focal adhesion of the cells to the culture vessel. EDTA, which is often added together with trypsin, acts as an important chelating agent to remove Ca and Mg, both of which are important in the maintenance of tight junctions, such as cadherins and integrins. Combined together, trypsin/EDTA works by cleaving cell-vessel and cell-cell adhesion.
Some trypsin/EDTA alternatives include:
- Papain - for sensitive cells (e.g. neurons)
- Collagenase/Accutase - for trypsin-insensitive cells
- PBS/EDTA - might take significantly longer for the cells to detach and some knocking off action might be required to initiate detachment.
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