I have read many user manuals but I could'nt find some specific data for cells in suspension as EBV immortalized B lymphocytes... And this is a culture for cell production not for protein production...
For production of EBV virus from suspended cells, the producer cells are seeded at 100,000 cells per ml in RPMI medium with 10% FCS. The rolling bottle is filled to 200ml and its rotation speed is 1 turn for 4 minutes. The harvesting speed is 4 days. For a line or clone B cells transformed by EBV, it is substantially the same thing but the culture medium is richer: DMEM / F12 supplemented with culture additive (Caryoser or ADCM or booster reagent). The harvesting speed is then 7 days. Finally for the development of an LLC, the medium should very rich linoléa supplemented with albumin / insulin / transferrin. The monitoring is done after 2 days of culture because the phenotype of these cells is very variable depending on the cell density and the depletion of the culture medium.
Thank you very much for all these precious informations... My problem now is that the Thermo roller I will use is indicated "1 to 80rpm speed"... We'll see...
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