BILODEAU: As I mentioned earlier, there’s good translation from flat-stock cultureware to the CellCube because it’s the same growth surface material— treated polystyrene. There will be some necessary troubleshooting and optimization. The better you know your cells and the more you’ve characterized your cultures, the smoother that transition is going to be.
Specifically, it’s important to understand the timing of cell attachment and cell spreading in static culture before you move into the CellCube. It’s also helpful to understand the components of your medium and how they might change during cell growth. For instance, are the cells using a lot of glucose so that you can use glucose consumption as an indicator of expansion? Typically, we recommend starting with a CellCube 10- or 25-layer module, seeding with the standard cell density and medium that you use for maintenance cultures for your cell line. You plan for a typical expansion period and then start with middle-of-the-road medium conditioning. That would be a pH of 7.35 to 7.4 and 50% dissolved oxygen unless you have data to support different parameters. Monitor nutrient metabolite concentration offline regularly during the cell expansion. Harvest with your preferred association reagent and see what your yield is. Then, you would adjust the seeding, medium volume, medium-conditioning parameters to reach your desired yield per cm2 on that small scale.