Cell passaging
Adherent cultures should be passaged when they are in the log phase before they reach confluence, otherwise, it takes them longer to recover when reseeded. Normally, you can detect that visually by estimating the percentage of confluence as the amount of free space between cells. Usually, the normal range to perform the cell passage is
Disassociation techniques
The first step in subculturing adherent cells is to detach them from the surface of the culture vessel. You can do that using enzymatic or mechanical procedures, as you can see in the table below.
Procedure | Dissociation agent |
---|---|
Shake-off | Shaking or rocking of culture vessel, or vigorous pipetting |
Scraping | Cell scraper |
Enzymatic dissociation | Trypsin, Trypsin+collagenase, Dispase, TrypLETM |