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A technique for increasing yields in bioreactors and disposable cell-culture systems


FibraCel disks are a new cell-growth matrix that dramatically increase yields in packed-bed reactors, and have also proven highly effective when used in spinners, disposable bottles and bags. New Brunswick Scientific offers two new devices that use FibraCel disks to increase productivity: the FibraStage, a simple and disposable culture system that outperforms spinners, T-flasks and roller bottles, and the CelliGen 310, an advanced benchtop bioreactor suitable for research and development or small-scale production.


What is FibraCel?

FibraCel disks are a patented, solid-support matrix for growth of both anchorage-dependent and suspension cultures. Designed for the growth of animal and insect cells (Table 1), they are predominantly used for production of secreted products such as recombinant proteins and viruses. FibraCel disks were introduced in the early 1990s, and today are used throughout the world for both research and production, including the manufacture of the worlds first licensed gene-therapy drug.

Manufactured according to cGMP guidelines, FibraCel disks are composed of a polyester nonwoven fiber bonded to polypropylene mesh, providing a three-dimensional substrate to which cells can readily attach or in which they become entrapped (Fig. 1).

In a packed-bed bioreactor, the FibraCel disk bed serves to immobilize the cells, providing a low-shear, low-foam environment, and an extremely high-surface area for cell growth. Cells remain entrapped in the disk bed throughout the run, leaving the medium cell-free for simplified medium replacement and product collection 1 . Exceptionally high yields, typically 108 cells/cm3 of bed volume depending on cell line, have been reported2 (Table 2). Foaming is virtually eliminated. Furthermore, cells generally attach within 1560 min on FibraCel, compared to approximately six hours for microcarriers. FibraCel disks are inoculated in a single step. The disks are autoclavable and, unlike microcarrier beads, do not fracture or lose their integrity through use.


CELL BIOLOGY CELL CULTURE APPARATUS


A new labor-saving, disposable culture device
An inexpensive new cell-culture system using FibraCel disks in disposable bottles is now available. FibraStage bottles are presterilized and preloaded with 10 g of FibraCel disks, into which users can inoculate cells with 500 ml of medium. Gas exchange and medium mixing are accomplished simply and efficiently by placing up to four bottles in a FibraStage device, which slowly raises or lowers the medium level in the bottles by collapsing or expanding the bellows at the base of each bottle. The entire device can be placed inside of a standard CO2 incubator for temperature and gas control.

The system is capable of producing milligram quantities of proteins and virus as well as biomass from a wide variety of cell types.

A single FibraStage bottle can replace the need to maintain dozens of roller bottles and hundreds of T-flasks. Because each bottle works as an independent reactor, you can easily use this as a screening device to test different medium formulations or various cell lines. In the US and UK, free 30-day in-lab trials are offered. For details, see http://www.nbsc.com/n2.htm.


CelliGen 310 bioreactor for advanced processing
At the opposite end of the control spectrum is New Brunswick Scientifics sophisticated new autoclavable benchtop unit, the CelliGen 310. This next-generation bioreactor-fermentor provides built-in control of up to 32 process parameters per vessel, with the ability to control up to four reactor vessels, or over 120 control loops, simultaneously. Features include the ability to choose up to five thermal mass flow controllers for sparge and gas overlay, or connect more than 10 external devices, such as sensors, scales or gas analyzers, for optimized process control. A large, 15-inch touch-screen interface simplifies entering and viewing setpoints, alarms, trend graphs and recipes.

A variety of specialized cell culture vessels and impellers is available, including an optional packed-bed basket for applications requiring high yields of secreted products. The CelliGen 310 cell culture vessels are available in four sizes, ranging from 2.2 to 14.0 l total volume, and when used with the packed-bed basket, require 50500 g of FibraCel disks per run.

FibraCel offers the added benefit of sustaining long-term periods of high-density growth in perfusion. There is never any danger of clogging. Higher cell densities and productivities coupled with easier downstream processing also lower overall production costs. Elimination of cell filtration (including spin filters), lowers costs still further. Cells immobilized in the matrix serve as a renewable catalyst for continuous production of proteins. Fresh medium is fed into the bioreactor, washing secreted products through the matrix to provide a cell-free effluent. For additional information about this packed-bed bioreactor, see our website (http://www.nbsc.com/products/bioreactors/CelliGenPlus2. asp).


1. Kaufman, J.B. et al . Continuous production and recovery of Ca2+ binding receptor from HEK 293 cells using perfusion through a packed bed bioreactor. Cytotechnology 33, 311 (2000).

2. Wang, G. et al. Modified CelliGen-packed bed bioreactors for hybridoma cell cultures. Cytotechnology 9, 4149 (1992).


Rajul Vyas & Julia Cino
New Brunswick Scientific, PO Box 4005, Edison, New Jersey 08818, USA. Correspondence should be addressed to R.V. (bioinfo@nbsc.com).


This article was submitted to Nature Methods by a commercial organization and has not been peer reviewed. Nature Methods takes no responsibility for the accuracy or otherwise of the information provided.



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