Nunc cell factories

Fig. 2.3. FPLC purification of rAAV. Ten Nunc cell factories of rAAV-UFll...

Fig. 3.4. A Nunc cell factory. (Courtesy of Nunc and Gibco.)...

Growth of hybridoma cells in Nunc Cell Factories... 

Growth of hybridoma cells in stationary monolayer cultures 130 Growth of hybridoma cells in Nunc Cell Factories 131... 

On the other hand, the principle of CellCube and Cell Factory is to provide a large area for cell adhesion and proliferation in relatively compact devices. The latter consists of multiple trays, providing surface areas in the range of 700-25 000 cm2, for medium volumes of 0.2-8 L (Nunc, 2006). The system called CellCube provides a surface area in the range of 8500-85 000 cm2 and is fitted with an automated control system (Corning, 2006). 

The roller bottle technique is a well-established and successful culture method widely used for the production of cells and products. One reason for this is that a single contamination event does not mean that the whole batch is lost, as with a single unit process. However, roller culture needs a considerable financial investment both in the apparatus itself and in incubator/hot-room facilities. Also, some cell lines (particularly epithehal) may not be as successfully grown in roller bottles as in stationary bottles. Common problems are streaking, clumping or inadequate spreading over the total surface (e.g. non-locomotory cell lines). An alternative scale-up route is to use multisurface stationary systems such as the Nunclon Cell Factory (Nunc) (see section 5.7) or the CellCube (Costar). 

Seed [10] ten-layer cell factories (Nunc) each with 5x10 293 cells so that each will be 75-80% confluent the next day. (Incubate 12-16h prior to transfection.) 293 cells are cultured in DMEM supplemented with 5% fetal bovine serum, 1% penicillin, and 1% streptomycin. 

An extension of the flask culture principle for giving large unit surfece areas are the Nunc A/S Cell Factories (16). These are polystyrene plates of 632 cm which are available in single, double (1264 cm ), 10 tray (6320 cm ), and 40 tray (25280 cm ) units. Each tray level requires 200 ml medium and can be filled, harvested, etc. in one operation per fectoiy unit. For large scale commercial processes the system can be semi-automated with the Manipulator and Robot System. 

Cell Factory (A/S NUNC Roskilde, Denmark) The trays are 335 x 205 mm (600 cm ) and can be obtained in multiples of 1, 2, 10, and 40. The largest has a surface area of 24,000 cm and requires 8 L medium (equivalent to 14 large roller bottles but requiring half the incubation space and no ancillary roUer equipment). 

Cell seed expansion A series of culture steps is needed to expand the cell seed ampoule (e.g., 5 million cells) to production size (range 10 to 10 cells). For HDC, this is accomplished in steps of a split level of 1 2 or more usually 1 4 through a series of flasks, roller bottles, and possibly cell factories (A/S Nunc). Other cell types are split at a 1 5 to 1 20 ratio. A similar build-up is needed for the virus seed. 

Figure 1.7 (a) The Nunc automatic cell factory manipulation (ACFM) system of Thermo Fisher Scientific Inc., a well-established system to grow cells on multiple flat bottoms. In order to reach the high cell numbers required in clinical applications, however, stirred disposable bioreactors are increasingly used to grow cells on... 

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