The CRL-3216 cells are particularly adapted to the ATCC-formulated DMEM, catalog no. 30-2002.
ATCC DMEM (30-2002), is formulated with 1.5 g/L sodium bicarbonate for use in 5% CO2 incubators. Other DMEM is formulated with 3.7 g/L sodium bicarbonate for use in 10% CO2. When using medium with high bicarbonate these cells must be incubated in 10% CO2 so that the medium is buffered correctly. When the medium is not buffered properly these cells will not be able to adhere and will remain floating in the medium, although viable.
Also be sure to add the extra 2m M L-glutamine to the formulation.
When grown in the recommended ATCC medium, CRL-3216 cells are typically healthy and well-attached with an epithelial-like morphology. A culture of CRL-3216 initiated in a T75 flask with 15ml of complete growth medium will be ready for subculture within a few days. When refreshing the medium during the first few days post thaw, retain any floating cells by gentle centrifugation and add them back to the flask with the adherent cells. Discarding viable floating cells can make the culture too dilute and growth will lag. Also, if these cells become over-confluent they may detach from the flask, retain these floating cells and subculture using 0.05% Trypsin-0.53 mM EDTA (Gibco 25300).
|Date Created||01/30/2018 03:19 PM
|Date Updated||01/30/2018 03:23 PM