One-Step Seed Culture Expansion from One Vial of a High-Density Cell Bank to a 2,000-L Production Bioreactor

We describe the use of perfusion cell culturing to reduce processing time, simplify operations, and maximize equipment use in seed culture expansion processes. Seed culture expansion is commonly performed using several consecutive batch cultures. Starting from a cryopreserved cell stock, the initial culture expansion is typically performed using shake flasks, whereas the final steps are bioreactor cultures. Such a procedure is time consuming and labor intensive, and each manual interaction constitutes a contamination risk for the culture. Here, perfusion was used to simplify preparation of a high-density cell bank without the need for a separate centrifugation step. In combination with perfusion, this cell bank was used to shorten the overall seed expansion process, from cryovial to the N – 1 bioreactor step (Figure 1).

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