Maintenance of the cells on day 1, 3 (and 6) Catalog Number HPR116

Note: The users have two options: Either use the cells immediately after thawing, or following at least 3 days of culture (best 7 days). HepaRG® cells keep a high level of CYP activities during the first 24 hours following thaw and plating, and these activities then decrease while the cells reconstitute the monolayer, then the activities return during the fourth day in culture, peaking at Day 8.

1. To maintain the cells, change the media on day 1, 3 (and 6).
2. Before every media change, take pictures of the cells in order to follow their proper attachment and restoration of function. Always follow the same pattern (e.g. 2B, 4D, 5E etc).
3. Prepare the maintenance media by adding the HepaRG® Maintenance and Metabolism Medium 620 supplement (ADD620 BPI) to the bottle of Basal Hepatic Medium (MIL600 BPI). Once ready, the media can be stored at +4 oC for a period of one month.
4. Pre-warm (37°C) the HepaRG® Maintenance and Metabolism Medium 620.
5. Aspirate the medium.
6. Gently add the pre-warmed HepaRG® Maintenance and Metabolism Medium 620 to the side of each well with an automatic single channel pipette (100 µL/well for a 96 multi-well plate). Do not add the medium directly onto the cells!

Depending on the scientific question several media options are available for the actual intended application.