Roller bottle systems

The roller bottle system is a multiple process requiring considerable staff time for repeated manipulations. Thus many modifications have been introduced that increase the efficiency or surface area capacity of roller bottles. Some examples are as follows ... 

A simple way of culturing anchorage-dependent animal cells, and stem cells in particular, under dynamic conditions is provided by roller bottles systems (Kunitake et al., 1997 Mitaka, 2002). Cell culture in roller bottles consists of placing multiple cylindrical bottles (250-2000 mL) into an apparatus that will rotate the bottles at rotational speeds of 5-60 rph (ECACC/Sigma-Aldrich, 2010). The entire internal surface of the bottle is used for cell growth. With this system, the cells are more efficiently oxygenated due to alternative exposure to the medium and the gas phase. Apparatus are available to... 

Different animal cell types display different properties pertinent to their successful culture. Those used to manufacture biopharmaceuticals are invariably continuous (transformed) cell lines. Such cells will grow relatively vigorously and easily in submerged culture systems, be they roller bottle or bioreactor based. 

Automated roller bottle cell culture systems, 5 35 It... 

Commercial scale cultivation of mammalian cells is accompHshed using different technologies roller bottles, microcarriers, suspension (batch, fed-batch or perfusion mode) and hollow fiber bioreactors (Table 2). However, especially for products needed in large amounts, suspension cultivation seems to be the most effective system [4, 5]. Suspension-based systems are characterized by a homogeneous concentration of cells, nutrients, metabolites and product, thereby facilitating scale-up [6] and enabling an accurate monitoring and control of the culture [7]. 

Stirred-tank bioreactors Air-lift bioreactors Wave bioreactors Microcarrier-based systems Packed-bed bioreactors Fluidized-bed bioreactors Hollow-fiber bioreactors Bioreactors providing surfaces for attached cell growth (roller bottles, CellCube , Cell Factory)... 

The basic need for a solid support guides all production choices involving industrial processes for adherent cells. A large variety of vessels has been developed for adherent cell cultures. Petri dishes, Roux bottles, T-flasks, and roller bottles are examples of cell culture vessels with a glass or polystyrene surface. The system of choice is dependent on the seal-ability of multiple steps, as well as the cost of equipment and qualified operators. 

Alternatively, some subunit viral vaccines can be generated by rDNA techniques and expressed in a continuous cell line or insect cells. Recent advances in bio reactor design and operation have improved the successful production of IPV in large-scale bioreactors. However, roller bottles or flasks are still used for most current vaccine production. Development of insect cell culture will allow for very large-scale liquid suspension culture (143). Several vaccine candidates such as gpl60 for HIV and gD protein for herpes have been demonstrated in the insect cell culture system. However, no vaccine has... 

Bellco-Corbeil Culture System (Bellco). A roller bottle is packed with a cluster of small glass tubes (arranged in parallel and separated by silicone spacer rings). Models are available in lOOO-cm, 10 OOO-cm and 15 000-cm sizes. Medium is perfused through the vessel and the bottles rotate 360° in alternate directions to avoid tube twisting and the need for special caps. 

If hundreds of roller bottles need to be handled, then a robotic culture system (Cellmate, The Automation Partnership, Royston, UK) could be considered, which reduces manning by 10-fold as well as giving improved consistency and reproducibility. 

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). 

Suspension culture systems, unlike bottles, etc., can be environmentally controlled and were optimized in the late 1960s to a minimum 100-1 capacity, i.e. a 1 X100-1 fermenter was equivalent to 800-1500 roller bottles. The scale-up potential is 10 0001 (for suspension cells), although currently only 40001 has been used for microcarrier culture. 

For detailed cost analysis, a 10-L roller bottle crdture, a conventional stirred-tank reactor as well as a Wave system were... 

Integration of fixed costs, consumables and manpower requirements on a typical industry cost basis showed that the Wave " system was far the most economical solution ( 1297 per run), while roller bottles ( 1789 per ran) and the stirred-tank reactor ( 2224 per ran) were up to 70% more cost-intensive (Fig. 14.3, Table 14.7). 

System Roller bottles Stirred-tank reactor Wave bioreactor... 

Production The production culture may be a batch of several hundred roller bottles, 30 to 50 cell factories, or a single bioreactor for suspension (100 to 10,000 L) or microcarrier (50 to 500 L) ceUs. Although batch-type production is still the most common process, continuous processes where the product is harvested daily over a long period (20 to 100 d) are being increasingly used. Culture systems based on hollow fibers, porous microcarriers, or other immobilization techniques are used for continuous perfusion processes. During the production phase, the virus seed or a promoter (e.g., for interferon) may be added. 

The steps between the cell bank and the production fermentation serve the purpose of expanding the seed volume (biomass) to finally provide sufficient cells to inoculate the main fermenter. Early seed stages, also referred to as precultures, are often run in shake flasks or in T-flasks and roller bottles in the case of cell culture, whereas subsequent stages are performed in stainless steel or single-use bioreactors. The dimensions of the seed bioreactors are adjusted to the biological system, as shown in Table 1.5. 

Roller bottles Simple operation and usage Versatile system Low-cost solution Monitoring and control is possible, but not straightforward Although minimized, concentration gradients still persist... 

Kunitake R, Suzuki A, Ichihashi H, Matsuda S, Hirai O, Morimoto K. 1997. Fully-automated roller bottle handhng system for large scale culture of mammalian cells. / Biotechnol 52(3) 289-94. 

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